The document discusses various methods of automation used in microbiology laboratories, including automation of specimen collection, transport, processing, microscopy, identification and antimicrobial susceptibility testing. It describes several automated systems that have been developed to perform functions like automated staining, urine sediment analysis, specimen plating, bacterial identification using techniques like MALDI-TOF, VITEK and reaction-based systems. The document also provides examples of various automated equipment and their applications in microbiology laboratories.
The document summarizes various automated systems used in bacteriology. It discusses automated blood culture systems like BacT/ALERT 3D, BD BACTEC, and VersaTREK which continuously monitor blood cultures. It also describes automated identification and antimicrobial susceptibility testing systems like Vitek, Phoenix, and MicroScan Walkaway. Finally, it provides an overview of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) which uses the mass spectra of proteins to rapidly identify bacteria and yeast.
This document discusses various methods for automated detection and identification of microorganisms in clinical microbiology laboratories. It describes several automated blood culture systems that use instrumental devices to incubate and monitor blood culture bottles. It also discusses rapid identification systems like API 20E strips and instruments like Vitek 2, BD Phoenix, and Microscan Walkaway that can identify bacteria based on biochemical testing of microorganisms. The document concludes with an overview of MALDI-TOF mass spectrometry which enables rapid protein-based identification of bacteria and fungi from cultures.
This document discusses automation in microbiology labs. It outlines several technologies currently used for semi-automation, including blood culture incubators, ID and susceptibility testing platforms, and automated stainers. Fully automated total lab automation (TLA) systems are also described, which use conveyor systems and digital imaging to automate specimen processing and plate reading. Advantages of TLA include continuous incubation, flexibility in plate reading times, efficient retrieval of plates as needed, and improved traceability. However, more research is still needed to assess the benefits of full microbiology automation.
Automation provides benefits to clinical microbiology such as continuous monitoring of blood culture systems to detect pathogens faster. Automated identification and susceptibility testing systems like Vitek-2 can analyze test results every 15 minutes using optical detection of signals. Molecular diagnostic techniques allow for identification of pathogen nucleic acids.
The use of a machine designed to follow repeatedly and automatically a predetermined sequence of individual operations.
AUTOMATED WASHING
AUTOMATED MEDIA PREPARATORS
AUTOMATED COLLECTION AND
PROCESSING OF SAMPLES
CYTOSPIN
AUTOMATED GRAM STAINING
AUTOMATED STREAKING
SPIRAL PLATER
AUTOMATED ANTIBIOTIC -
SENSITIVITY SYSTEM
AUTOMATIC COLONY COUNTER
AUTOMATED URINE MICROSCOPY -
ANALYSER
Automation in microbiology, changing concept and defeating challengesAyman Allam
This document discusses automation in microbiology and some of the challenges associated with it. It begins by providing background on the early history of microbiology. It then discusses how rapid and accurate microbiological results are important for patient care. Automation provides advantages like rapid results, reproducibility, and reduced errors. However, microbiology is a complex field that still requires human input for interpreting results. The document reviews several automated systems for blood culture, identification, and antibiotic susceptibility testing. It also evaluates one such system, the Phoenix system, and finds it provides generally good identification and AST results compared to conventional methods. In conclusion, automation can be suitable for high-volume laboratories but cost must be considered, as microbiology still requires human expertise.
Automated system for bacterial identificationDEEKSHANT KUMAR
[DOWNLOAD IT OPEN IT WITH MICROSOFT POWERPOINT THEN YOU WILL BE ABLE TO UNDERSTAND THE TOPIC COVERED.]
1. WHOLE TEXT IS RELIABLE.
2. TEXT HAS BEEN TAKEN FROM STANDARD TEXT BOOK FOR MEDICAL MICROBIOLOGY.
3. SOME PICTURE HAS BEEN TAKEN FROM JOURNAL.
This document discusses antibiotic susceptibility testing for aerobic bacteria. It describes various antibiotic classes and provides examples of antibiotics within each class. It also discusses new antibiotics for treating multi-drug resistant gram-negative bacteria. The major mechanisms of antibiotic resistance are outlined. Finally, it summarizes methods for susceptibility testing and discusses antibiotic resistance in important bacteria such as MRSA, VRE, ESBL producers, and carbapenemase producers.
The document summarizes various automated systems used in bacteriology. It discusses automated blood culture systems like BacT/ALERT 3D, BD BACTEC, and VersaTREK which continuously monitor blood cultures. It also describes automated identification and antimicrobial susceptibility testing systems like Vitek, Phoenix, and MicroScan Walkaway. Finally, it provides an overview of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) which uses the mass spectra of proteins to rapidly identify bacteria and yeast.
This document discusses various methods for automated detection and identification of microorganisms in clinical microbiology laboratories. It describes several automated blood culture systems that use instrumental devices to incubate and monitor blood culture bottles. It also discusses rapid identification systems like API 20E strips and instruments like Vitek 2, BD Phoenix, and Microscan Walkaway that can identify bacteria based on biochemical testing of microorganisms. The document concludes with an overview of MALDI-TOF mass spectrometry which enables rapid protein-based identification of bacteria and fungi from cultures.
This document discusses automation in microbiology labs. It outlines several technologies currently used for semi-automation, including blood culture incubators, ID and susceptibility testing platforms, and automated stainers. Fully automated total lab automation (TLA) systems are also described, which use conveyor systems and digital imaging to automate specimen processing and plate reading. Advantages of TLA include continuous incubation, flexibility in plate reading times, efficient retrieval of plates as needed, and improved traceability. However, more research is still needed to assess the benefits of full microbiology automation.
Automation provides benefits to clinical microbiology such as continuous monitoring of blood culture systems to detect pathogens faster. Automated identification and susceptibility testing systems like Vitek-2 can analyze test results every 15 minutes using optical detection of signals. Molecular diagnostic techniques allow for identification of pathogen nucleic acids.
The use of a machine designed to follow repeatedly and automatically a predetermined sequence of individual operations.
AUTOMATED WASHING
AUTOMATED MEDIA PREPARATORS
AUTOMATED COLLECTION AND
PROCESSING OF SAMPLES
CYTOSPIN
AUTOMATED GRAM STAINING
AUTOMATED STREAKING
SPIRAL PLATER
AUTOMATED ANTIBIOTIC -
SENSITIVITY SYSTEM
AUTOMATIC COLONY COUNTER
AUTOMATED URINE MICROSCOPY -
ANALYSER
Automation in microbiology, changing concept and defeating challengesAyman Allam
This document discusses automation in microbiology and some of the challenges associated with it. It begins by providing background on the early history of microbiology. It then discusses how rapid and accurate microbiological results are important for patient care. Automation provides advantages like rapid results, reproducibility, and reduced errors. However, microbiology is a complex field that still requires human input for interpreting results. The document reviews several automated systems for blood culture, identification, and antibiotic susceptibility testing. It also evaluates one such system, the Phoenix system, and finds it provides generally good identification and AST results compared to conventional methods. In conclusion, automation can be suitable for high-volume laboratories but cost must be considered, as microbiology still requires human expertise.
Automated system for bacterial identificationDEEKSHANT KUMAR
[DOWNLOAD IT OPEN IT WITH MICROSOFT POWERPOINT THEN YOU WILL BE ABLE TO UNDERSTAND THE TOPIC COVERED.]
1. WHOLE TEXT IS RELIABLE.
2. TEXT HAS BEEN TAKEN FROM STANDARD TEXT BOOK FOR MEDICAL MICROBIOLOGY.
3. SOME PICTURE HAS BEEN TAKEN FROM JOURNAL.
This document discusses antibiotic susceptibility testing for aerobic bacteria. It describes various antibiotic classes and provides examples of antibiotics within each class. It also discusses new antibiotics for treating multi-drug resistant gram-negative bacteria. The major mechanisms of antibiotic resistance are outlined. Finally, it summarizes methods for susceptibility testing and discusses antibiotic resistance in important bacteria such as MRSA, VRE, ESBL producers, and carbapenemase producers.
The document discusses antimicrobial susceptibility testing (AST), which determines the susceptibility of bacteria to different antimicrobial agents through in-vitro laboratory procedures. It outlines various guidelines and standardization procedures for AST, including standardized bacterial inoculum, growth medium, incubation conditions, and antimicrobial concentrations. Common AST methods include dilution methods like broth microdilution and agar dilution, which determine minimum inhibitory concentrations (MICs), and diffusion methods like disc diffusion testing, which provide qualitative susceptibility results.
Blood culturing is the most important test for detecting pathogens in the bloodstream. It involves collecting blood in specialized bottles that contain growth media for aerobic and anaerobic organisms. It is critical that the collection procedure is done aseptically. Newer automated systems can continuously monitor blood cultures and detect microbial growth within 24-48 hours, providing faster results than conventional methods. Rapid identification of pathogens in positive blood cultures is important for guiding appropriate treatment.
The document discusses antibiotic sensitivity testing (AST), which measures the susceptibility of bacterial isolates to different antibiotics. AST aims to select the most appropriate antibiotic for patients and assess emerging resistance patterns. Common AST methods include disk diffusion, broth dilution, Etest, and automated systems. Disk diffusion is the most widely used method and involves placing antimicrobial disks on agar plated with the test organism and measuring inhibition zone sizes. Broth dilution determines the minimum inhibitory concentration (MIC) by visually inspecting bacterial growth in serial antibiotic dilutions.
The document discusses laboratory diagnosis of bacterial infections. It describes how automated systems can identify bacteria faster and more accurately than conventional methods. Several automated systems are highlighted, including BacT/ALERT and VITEK, which continuously monitor blood culture bottles for microbial growth. MALDI-TOF enables rapid bacterial identification by examining ribosomal protein patterns. VITEK 2 and other automated systems can identify bacteria and perform antimicrobial susceptibility testing in a single system within a day. Overall, the document outlines the benefits of automation for improving the efficiency and accuracy of bacterial diagnostics.
This document summarizes different systems for processing blood cultures. It describes manual and automated blood culture systems, as well as various broths, additives, and detection methods. Key automated systems discussed include BacT/Alert, BACTEC, and Isolater. BacT/Alert is described in more detail, outlining its media, bottles with CO2 sensors, colorimetric detectors, and computerized analysis to detect microbial growth based on changes in CO2 concentration over time.
Quality control of antimicrobial susceptibility testsAmr Eldakroury
This document discusses quality control for antimicrobial susceptibility testing. It describes different testing methods, including disk diffusion and microdilution, and how they provide qualitative or quantitative results. Key aspects of quality control are highlighted, such as using standardized reference strains, maintaining proper test conditions for media, antimicrobials, inoculum, incubation, and ensuring accurate interpretation. Close monitoring is important to identify potential errors at each step and ensure accurate patient results.
This document provides an overview of quality assurance and quality control in microbiology. It discusses the importance of quality in microbiology laboratories given the consequences of unreliable test results. It defines key terms like quality assurance and quality control and describes various quality concepts. It also outlines factors that can influence quality like pre-analytical, analytical and post-analytical factors. The document discusses organization of microbiology laboratories, external and internal quality assessment, and quality control of different laboratory sections, equipment, and preservation of stock cultures.
This document discusses quality assurance and quality control procedures for microlaboratories. It defines quality and quality management, and explains how quality is ensured in laboratories through controlling all factors that influence reliable test results. Quality assurance aims to ensure accurate and reliable data generation and use. Key quality control procedures discussed include regular equipment maintenance and calibration, sterilization validation methods, reagent and media quality control, and environmental monitoring. Adhering to these standardized quality control protocols is important for generating credible results and safeguarding patient health.
This document discusses antifungal susceptibility testing. It provides background on the history of antifungal susceptibility testing and why it is needed. It describes different methods for testing including broth dilution and disk diffusion. It discusses various antifungal agents and their mechanisms of action. The document outlines the procedures for broth microdilution and macrodilution testing according to CLSI guidelines, including preparation of inoculum, drug solutions, reading results, and testing of filamentous fungi.
This document discusses bacteriocin typing for epidemiological investigations. It defines bacteriocins as bactericidal proteins produced by bacteria that kill closely related bacterial strains. Bacteriocin typing involves determining the bacteriocin production patterns of strains against indicator strains or testing strains for susceptibility to different bacteriocins. This allows differentiation of bacterial isolates and investigation of outbreaks. Specific examples discussed are colicin typing of E. coli and pyocin typing of P. aeruginosa. The document outlines the methods for these typing techniques.
Automated blood culture systems like BacT/ALERT and BACTEC provide continuous monitoring of blood culture specimens to more quickly detect pathogens. They work by monitoring changes in carbon dioxide or fluorescence levels that occur as pathogens metabolize nutrients in the culture bottles. This allows for earlier detection compared to conventional manual methods. Popular systems include BacT/ALERT, BACTEC, Vital, and VersaTREK systems. They have increased pathogen detection rates while reducing the hands-on time needed compared to older techniques.
This document discusses quality control in microbiology. It covers various aspects of quality control including the pre-analytic, analytic and post-analytic phases of diagnostic testing. In the pre-analytic phase, it discusses proper specimen collection, transport and processing. The analytic phase covers microscopic examination, culture processing, identification and antimicrobial susceptibility testing. It also discusses quality control of equipment, reagents and culture media. The post-analytic phase involves reporting of results and interaction with epidemiologists. Maintaining accurate records and samples is also emphasized.
This document discusses sensitivity testing, which determines the effectiveness of antimicrobial drugs against bacterial strains. Sensitivity testing helps select the most effective drug and identify pathogens. It is done using diffusion or dilution techniques to measure zones of inhibition or minimum inhibitory concentrations. However, sensitivity testing only measures drug effectiveness in laboratory conditions and does not account for patient factors. Antimicrobial resistance can develop through genetic mutations or exchange between bacteria, allowing resistance to spread to multiple drugs. Sensitivity testing guides optimal treatment while addressing the rise of antimicrobial resistance.
The document discusses the inducible clindamycin resistance test (D test) which is recommended to test for inducible resistance in staphylococci, streptococcus pneumoniae, and beta-hemolytic streptococci that are erythromycin resistant but clindamycin susceptible. The D test detects resistance by observing whether the zone of inhibition around a clindamycin disk is flattened near an erythromycin disk. A positive result indicates inducible resistance and that clindamycin treatment may fail in vivo due to selection of resistant mutants. The procedure and interpretation of the D test is described along with its limitations and clinical significance when evaluating clindamycin susceptibility.
Antibiotic Sensitivity Testing 2020 Update Margie Morgan
This document discusses antibiotic sensitivity testing and provides information on:
1. Common antibiotic classes and mechanisms of resistance.
2. Methods for antibiotic sensitivity testing including disk diffusion, E test, and broth dilution.
3. Important resistant bacteria like MRSA, VRE, ESBLs, CREs, and guidelines for detecting them.
4. Interpreting results and quality control for antibiotic sensitivity testing.
This document provides information about anti-fungal susceptibility testing. It discusses various fungi and the available anti-fungal drugs that act on different fungal targets like the cell wall, cell membrane, microtubules, RNA/DNA, and protein synthesis. It covers different testing methods like macrodilution, microdilution, and disk diffusion. It provides details on test medium, inoculum preparation, drug dilutions, and incubation conditions. It also discusses interpretive criteria, quality control strains and ranges, emerging resistance, and the need for susceptibility testing.
The document discusses good practices for collecting microbiology samples to obtain accurate diagnostic results. It emphasizes that sample quality depends on proper collection, transport, and clinical information provided. Specific guidelines are given for collecting various sample types like blood, respiratory, body fluids, wounds, and urine. Factors like aseptic technique, appropriate volume, and timely transportation are important for maximizing recovery of microorganisms.
This document discusses several instruments and techniques used in microbiology laboratories, including:
1. The Polystainer 5300, an automated system that can stain up to 20 slides in 5-10 minutes to aid rapid diagnosis.
2. The BACTEC 9240, an automated blood culture system that can detect bacteria in blood samples within days using sensors to detect changes in oxygen and carbon dioxide levels in culture vials.
3. The Vitek 2, an automated system that uses test cards containing 64 wells with metabolic substrates to identify microbes based on their reactions over 24-48 hours.
This document discusses various laboratory techniques for rapidly diagnosing and identifying microbes. It begins by explaining the need for rapid diagnosis and then describes several identification methods including biochemical profiling, API strips, BD BBL crystals, Vitek systems, Biolog, and MIDI Sherlock. These methods vary in their automation, databases, turnaround times, and whether they require gram stains but all aim to more quickly and accurately identify unknown microbes. The document concludes that automated systems have made analysis more reliable and reduced time and labor compared to older techniques.
The document discusses antimicrobial susceptibility testing (AST), which determines the susceptibility of bacteria to different antimicrobial agents through in-vitro laboratory procedures. It outlines various guidelines and standardization procedures for AST, including standardized bacterial inoculum, growth medium, incubation conditions, and antimicrobial concentrations. Common AST methods include dilution methods like broth microdilution and agar dilution, which determine minimum inhibitory concentrations (MICs), and diffusion methods like disc diffusion testing, which provide qualitative susceptibility results.
Blood culturing is the most important test for detecting pathogens in the bloodstream. It involves collecting blood in specialized bottles that contain growth media for aerobic and anaerobic organisms. It is critical that the collection procedure is done aseptically. Newer automated systems can continuously monitor blood cultures and detect microbial growth within 24-48 hours, providing faster results than conventional methods. Rapid identification of pathogens in positive blood cultures is important for guiding appropriate treatment.
The document discusses antibiotic sensitivity testing (AST), which measures the susceptibility of bacterial isolates to different antibiotics. AST aims to select the most appropriate antibiotic for patients and assess emerging resistance patterns. Common AST methods include disk diffusion, broth dilution, Etest, and automated systems. Disk diffusion is the most widely used method and involves placing antimicrobial disks on agar plated with the test organism and measuring inhibition zone sizes. Broth dilution determines the minimum inhibitory concentration (MIC) by visually inspecting bacterial growth in serial antibiotic dilutions.
The document discusses laboratory diagnosis of bacterial infections. It describes how automated systems can identify bacteria faster and more accurately than conventional methods. Several automated systems are highlighted, including BacT/ALERT and VITEK, which continuously monitor blood culture bottles for microbial growth. MALDI-TOF enables rapid bacterial identification by examining ribosomal protein patterns. VITEK 2 and other automated systems can identify bacteria and perform antimicrobial susceptibility testing in a single system within a day. Overall, the document outlines the benefits of automation for improving the efficiency and accuracy of bacterial diagnostics.
This document summarizes different systems for processing blood cultures. It describes manual and automated blood culture systems, as well as various broths, additives, and detection methods. Key automated systems discussed include BacT/Alert, BACTEC, and Isolater. BacT/Alert is described in more detail, outlining its media, bottles with CO2 sensors, colorimetric detectors, and computerized analysis to detect microbial growth based on changes in CO2 concentration over time.
Quality control of antimicrobial susceptibility testsAmr Eldakroury
This document discusses quality control for antimicrobial susceptibility testing. It describes different testing methods, including disk diffusion and microdilution, and how they provide qualitative or quantitative results. Key aspects of quality control are highlighted, such as using standardized reference strains, maintaining proper test conditions for media, antimicrobials, inoculum, incubation, and ensuring accurate interpretation. Close monitoring is important to identify potential errors at each step and ensure accurate patient results.
This document provides an overview of quality assurance and quality control in microbiology. It discusses the importance of quality in microbiology laboratories given the consequences of unreliable test results. It defines key terms like quality assurance and quality control and describes various quality concepts. It also outlines factors that can influence quality like pre-analytical, analytical and post-analytical factors. The document discusses organization of microbiology laboratories, external and internal quality assessment, and quality control of different laboratory sections, equipment, and preservation of stock cultures.
This document discusses quality assurance and quality control procedures for microlaboratories. It defines quality and quality management, and explains how quality is ensured in laboratories through controlling all factors that influence reliable test results. Quality assurance aims to ensure accurate and reliable data generation and use. Key quality control procedures discussed include regular equipment maintenance and calibration, sterilization validation methods, reagent and media quality control, and environmental monitoring. Adhering to these standardized quality control protocols is important for generating credible results and safeguarding patient health.
This document discusses antifungal susceptibility testing. It provides background on the history of antifungal susceptibility testing and why it is needed. It describes different methods for testing including broth dilution and disk diffusion. It discusses various antifungal agents and their mechanisms of action. The document outlines the procedures for broth microdilution and macrodilution testing according to CLSI guidelines, including preparation of inoculum, drug solutions, reading results, and testing of filamentous fungi.
This document discusses bacteriocin typing for epidemiological investigations. It defines bacteriocins as bactericidal proteins produced by bacteria that kill closely related bacterial strains. Bacteriocin typing involves determining the bacteriocin production patterns of strains against indicator strains or testing strains for susceptibility to different bacteriocins. This allows differentiation of bacterial isolates and investigation of outbreaks. Specific examples discussed are colicin typing of E. coli and pyocin typing of P. aeruginosa. The document outlines the methods for these typing techniques.
Automated blood culture systems like BacT/ALERT and BACTEC provide continuous monitoring of blood culture specimens to more quickly detect pathogens. They work by monitoring changes in carbon dioxide or fluorescence levels that occur as pathogens metabolize nutrients in the culture bottles. This allows for earlier detection compared to conventional manual methods. Popular systems include BacT/ALERT, BACTEC, Vital, and VersaTREK systems. They have increased pathogen detection rates while reducing the hands-on time needed compared to older techniques.
This document discusses quality control in microbiology. It covers various aspects of quality control including the pre-analytic, analytic and post-analytic phases of diagnostic testing. In the pre-analytic phase, it discusses proper specimen collection, transport and processing. The analytic phase covers microscopic examination, culture processing, identification and antimicrobial susceptibility testing. It also discusses quality control of equipment, reagents and culture media. The post-analytic phase involves reporting of results and interaction with epidemiologists. Maintaining accurate records and samples is also emphasized.
This document discusses sensitivity testing, which determines the effectiveness of antimicrobial drugs against bacterial strains. Sensitivity testing helps select the most effective drug and identify pathogens. It is done using diffusion or dilution techniques to measure zones of inhibition or minimum inhibitory concentrations. However, sensitivity testing only measures drug effectiveness in laboratory conditions and does not account for patient factors. Antimicrobial resistance can develop through genetic mutations or exchange between bacteria, allowing resistance to spread to multiple drugs. Sensitivity testing guides optimal treatment while addressing the rise of antimicrobial resistance.
The document discusses the inducible clindamycin resistance test (D test) which is recommended to test for inducible resistance in staphylococci, streptococcus pneumoniae, and beta-hemolytic streptococci that are erythromycin resistant but clindamycin susceptible. The D test detects resistance by observing whether the zone of inhibition around a clindamycin disk is flattened near an erythromycin disk. A positive result indicates inducible resistance and that clindamycin treatment may fail in vivo due to selection of resistant mutants. The procedure and interpretation of the D test is described along with its limitations and clinical significance when evaluating clindamycin susceptibility.
Antibiotic Sensitivity Testing 2020 Update Margie Morgan
This document discusses antibiotic sensitivity testing and provides information on:
1. Common antibiotic classes and mechanisms of resistance.
2. Methods for antibiotic sensitivity testing including disk diffusion, E test, and broth dilution.
3. Important resistant bacteria like MRSA, VRE, ESBLs, CREs, and guidelines for detecting them.
4. Interpreting results and quality control for antibiotic sensitivity testing.
This document provides information about anti-fungal susceptibility testing. It discusses various fungi and the available anti-fungal drugs that act on different fungal targets like the cell wall, cell membrane, microtubules, RNA/DNA, and protein synthesis. It covers different testing methods like macrodilution, microdilution, and disk diffusion. It provides details on test medium, inoculum preparation, drug dilutions, and incubation conditions. It also discusses interpretive criteria, quality control strains and ranges, emerging resistance, and the need for susceptibility testing.
The document discusses good practices for collecting microbiology samples to obtain accurate diagnostic results. It emphasizes that sample quality depends on proper collection, transport, and clinical information provided. Specific guidelines are given for collecting various sample types like blood, respiratory, body fluids, wounds, and urine. Factors like aseptic technique, appropriate volume, and timely transportation are important for maximizing recovery of microorganisms.
This document discusses several instruments and techniques used in microbiology laboratories, including:
1. The Polystainer 5300, an automated system that can stain up to 20 slides in 5-10 minutes to aid rapid diagnosis.
2. The BACTEC 9240, an automated blood culture system that can detect bacteria in blood samples within days using sensors to detect changes in oxygen and carbon dioxide levels in culture vials.
3. The Vitek 2, an automated system that uses test cards containing 64 wells with metabolic substrates to identify microbes based on their reactions over 24-48 hours.
This document discusses various laboratory techniques for rapidly diagnosing and identifying microbes. It begins by explaining the need for rapid diagnosis and then describes several identification methods including biochemical profiling, API strips, BD BBL crystals, Vitek systems, Biolog, and MIDI Sherlock. These methods vary in their automation, databases, turnaround times, and whether they require gram stains but all aim to more quickly and accurately identify unknown microbes. The document concludes that automated systems have made analysis more reliable and reduced time and labor compared to older techniques.
This presentation in mainly focused of understanding of automation and its utility in cytopathology. It will be very usefull for postgraduate in pathology, cytopathologist and cytotechnicians.
This document discusses microscale bioprocessing, which involves producing commercially desired products at a very small scale. This speeds up product delivery, reduces costs, and increases consumer benefit. Two main formats are micro well systems and microfluidic systems. Microwell systems involve testing samples in small wells in plates, while microfluidic systems use channels to manipulate fluids in the micrometer range. These techniques are being applied to areas like bioprocess optimization, molecular biology procedures, diagnostics, drug discovery, and more. Microfluidics is revolutionizing fields and its market is projected to reach $6 billion by 2020. Future applications may involve nanofluidics which deals with even smaller nanometer scale fluids.
The ThinPrep 5000 processor is a fully automated system that can process up to 20 gynecological or cytology samples per batch. It utilizes proven ThinPrep technology to disperse, collect, and transfer cells from patient samples onto microscope slides in a controlled manner. The system ensures chain-of-custody by matching barcodes between each sample vial and slide to avoid mix-ups.
This document discusses various laboratory aspects of antimicrobial chemotherapy including reasons for testing antimicrobial susceptibility, methods for performing susceptibility tests like disc diffusion tests and MIC tests, detecting resistance mechanisms, and assaying antimicrobial serum levels. It describes techniques like Kirby-Bauer disc testing, Stokes comparative testing, BSAC standardized testing, tube dilution and E-test MIC methods, and automated methods like Vitek. The goals are to direct antibiotic treatment, monitor resistance trends, identify organisms, and ensure therapeutic versus toxic drug levels.
Protein microarrays allow the immobilization and detection of large numbers of proteins on small surfaces. Three key steps in the protein microarray workflow are printing, surface selection, and imaging. Optimizing the printing process is important to minimize contamination between samples. The Omnigrid and Microgrid systems can print contact-style onto 3D substrates with controls to reduce surface damage. Multiplexed protein microarrays on plates allow high-throughput screening by testing many samples in parallel. NovaRay imaging supports multiple array formats and wavelengths for detection. An example experiment showed specific and reproducible detection of target proteins in individual wells of a multiplexed plate with no carryover between wells.
Microbiological analysis of food products is the use of biological, biochemical, molecular or chemical methods for the detection, identification or enumeration of microorganisms in a material. Here some of the common methods have been described.
Expoquimia 2011: Forum Biotech - Javier AmayaExpoquimia
The document describes HexaScreen® HexaBatch, a miniaturized bioreactor system for animal cell culture screening. It consists of a plate with 6 independent mini bioreactors and a workstation. Key features include:
- Online monitoring of pH, dissolved oxygen, and cell density to obtain kinetic data
- Automated control of temperature, agitation, and aeration
- Reduced time and costs for screening compared to traditional methods like T-flasks or benchtop bioreactors due to parallel experiments and minimized manual labor
- Applications include cell line selection, medium optimization, process development, and toxicity testing.
Automation in biochemistry refers to using instruments to perform biochemical tests with minimal human involvement. Automated systems can perform many steps like sample handling, reagent addition, reaction incubation, and measurement that were previously done manually. The main types of automated analyzers are continuous flow analyzers, discrete autoanalyzers, and random access analyzers. Continuous flow analyzers pass samples and reagents sequentially through a single analytical pathway. Discrete autoanalyzers separate each sample and reagent in individual containers, allowing multiple tests per sample. Random access analyzers perform tests on batches of samples, selecting tests for each sample. Automated systems provide benefits like higher throughput, reduced variability, and less manual labor, but also have high initial costs.
Protein microarrays, or protein chips, allow high-throughput analysis of protein interactions and functions. There are two main types - analytical protein microarrays mostly use antibody detection, while functional protein microarrays study protein interactions and activity. Proteins are immobilized on surfaces like glass slides and probed with labels to measure binding. Applications include disease diagnostics, proteomics, analyzing protein networks and complexes, and developing new treatments.
Laser scanning cytometry and liquid based cytologyanaonline
Liquid based cytology (LBC) has been introduced to improve cervical screening. It uses a liquid preservative instead of smearing cells directly on a slide. This allows automated processing to disperse cells and transfer them evenly to a slide. LBC reduces sampling errors, improves cell preservation and slide quality compared to conventional smears. However, it requires specialized equipment and is more expensive. Laser scanning cytometry is a technique that can analyze individual cells from LBC samples. It provides quantitative measurements and complements flow cytometry by allowing analysis of adherent cells and solid tissues.
Diagnostic Medical Microbiology - Traditional and Modern approachChhaya Sawant
Updated version of Diagnostic Microbiology - Traditional and Modern approach. The presentation is an overview of conventional techniques still used in many laboratories and new technologies such as Molecular- and Protein-based testing
Rapid methods of detection of food borne pathogensAnchal
Rapid methods of detection of foodborne pathogens include biosensors, microscopic methods, immunological detection methods, and molecular detection methods. Biosensors can detect pathogens through metabolic patterns, phenotypic expression, nucleic acid analysis, and pathogen interaction with cells. Microscopic methods include direct epifluorescent filter technique, flow cytometry, and solid-phase cytometry. Immunological methods like lateral flow devices and ELISA use antibodies to detect pathogens. Molecular detection uses techniques like fluorescent in situ hybridization and polymerase chain reaction to detect pathogen DNA. These rapid methods aim to reduce detection time from days to hours compared to traditional culture methods.
Rapid methods of detection of foodborne pathogens include biosensors, microscopic methods, immunological detection methods, and molecular detection methods. Biosensors can detect pathogens through metabolic patterns, phenotypic expression, nucleic acid analysis, and pathogen interaction with cells. Microscopic methods include direct epifluorescent filter technique, flow cytometry, and solid-phase cytometry. Immunological methods like lateral flow devices and ELISA use antibodies to detect pathogens. Molecular detection uses techniques like fluorescent in situ hybridization and polymerase chain reaction to detect pathogen DNA. These rapid methods aim to reduce detection time from days to hours compared to traditional culture methods.
This document summarizes a lecture on microfluidics and their applications including lab-on-a-chip devices. Key topics discussed include microfluidic applications in areas like blood analysis, biochemical detection, chemical synthesis, and DNA sequencing. Common microfluidic materials like silicon, glass, and polymers are also summarized. Fabrication techniques for polymers include casting, hot embossing, and injection molding. Lab-on-a-chip offers advantages of low cost, small sample/reagent sizes, and minimized harmful byproducts compared to conventional laboratories.
This document summarizes a lecture on microfluidics and their applications including lab-on-a-chip devices. Key topics discussed include microfluidic applications in areas like blood analysis, biochemical detection, chemical synthesis, and DNA sequencing. Common microfluidic materials like silicon, glass, and polymers are also summarized. Fabrication techniques for polymers include casting, hot embossing, and injection molding. Lab-on-a-chip offers advantages of low cost, small sample/reagent sizes, and minimized harmful byproducts compared to conventional laboratories.
Capanna technique for chronic osteomyelitisVaisHali822687
The Capanna technique is used to reconstruct large bone defects using a combination of a massive bone allograft and vascularized fibular autograft. The massive allograft fills the bone gap, while the vascularized fibula provides a blood supply to promote healing. Over several months, the grafts integrate to restore bone structure, function, and stability. This technique is commonly used to treat bone loss from conditions like infection, tumor resection, and trauma.
Chronic osteomyelitis is difficult to treat and eradicate completely. It is characterized by infected dead bone within soft tissue with poor blood supply, making systemic antibiotics ineffective. Surgical debridement of infected bone and soft tissue is usually required along with long-term antibiotics. Eliminating dead space after debridement can be challenging and may require bone grafting, antibiotic beads, or flaps to fill gaps and promote healing.
This document compares hemiarthroplasty (HA) and proximal femoral nailing (PFN) for treating intertrochanteric femoral fractures (IFF) in the elderly. HA allows for earlier weight bearing and mobility, leading to faster recovery and better long-term joint function compared to PFN. HA reduces complications associated with immobility. While PFN has disadvantages like longer bed rest, HA is suggested for unstable fractures and in patients over 90 years old due to its benefits in reducing mortality, stress, and improving quality of life.
This document discusses viral hemorrhagic fever (VHF) and focuses on arboviral infections. It describes that VHFs are caused by viruses from three groups, including arboviruses, which are transmitted by arthropod vectors like mosquitoes and ticks. The document then details several arboviruses, their symptoms, locations, vectors and animal reservoirs. It specifically examines dengue virus, describing its structure, transmission from mosquitoes to humans, antibody response and laboratory diagnosis methods like NS1 antigen detection and antibody detection.
Complement system and its synthesis + activationVaisHali822687
Proteins normally found in serum in inactive form, but when activated they augment the immune responses.
Complements constitute about 5% of normal serum proteins.
Their level does not increase following either infection or vaccination.
There are four main stages in the activation of any of the complement pathways.
Initiation of the pathway
Formation of C3 convertase
Formation of C5 convertase
Formation of membrane attack complex (MAC)
Typhoidal and non typhoidal salmonella.pptxVaisHali822687
This document discusses typhoidal and non-typhoidal salmonella. It begins by introducing salmonella as a gram-negative bacteria and describes its classification. It then discusses the different serotypes of salmonella including S. Typhi, S. Paratyphi A, B, and C which cause enteric fever. The document summarizes the pathogenesis, clinical features, diagnosis, and treatment of typhoidal salmonellosis. It provides details on the various laboratory techniques used to identify and diagnose salmonella such as culture-based methods, biochemical testing, and serological tests.
The medial and lateral femoral circumflex arteries are the primary blood suppliers to the adult femoral head. The medial femoral circumflex artery courses behind the femoral neck and gives rise to the extracapsular arterial ring at the base of the neck. The lateral femoral circumflex artery courses along the anterior femoral neck. In children, the ligamentum teres artery also contributes significantly to femoral head blood supply, but its importance decreases with age as the medial femoral circumflex artery becomes the predominant supplier by age 8.
An intertrochanteric fracture occurs between the greater and lesser trochanters of the femur. It commonly affects elderly osteoporotic patients, usually women in their 80s, following a simple fall. X-rays are used to diagnose this extracapsular hip fracture. Treatment involves early internal fixation with devices like the sliding hip screw or intramedullary hip screw to allow early ambulation and prevent complications of non-operative management. Complications can include failure of fixation, malunion, or nonunion if reduction or implant positioning is inadequate.
Extracapsular hip fractures occur in the region between the greater and lesser trochanters of the femur, often extending to the subtrochanteric region. They are classified based on their location and stability, with unstable fractures involving comminution posing greater surgical challenges. Closed reduction and internal fixation with devices like the sliding hip screw are commonly used for treatment. Open reduction may be required for unstable patterns or when anatomical reduction cannot be achieved closed.
1) Recent advances in TB diagnosis include automated liquid culture systems like MGIT 960 and molecular diagnostic tests like Xpert MTB/RIF, which can detect TB and rifampin resistance in under 2 hours.
2) WHO recommends eight TB diagnostic tools including LED microscopy, liquid culture, rapid speciation strips, Xpert/MTB-RIF, urine LAM assay, LAMP, LPA, and SL-LPA to detect drug resistance.
3) Newer centralized high-throughput NAATs like RealTime MTB, FluoroType MTB, Cobas MTB, and Max MDR-TB run on automated platforms and can process hundreds of samples with high accuracy in
Hemiarthroplasty versus proximal femoral nailing VaisHali822687
This document compares hemiarthroplasty (HA) and proximal femoral nailing (PFN) for treating intertrochanteric femoral fractures in the elderly. HA was found to have advantages over PFN including earlier weight bearing, faster recovery from stress, and better long-term joint function. Specifically, HA patients could weight bear within 10 days compared to longer bed rest for PFN patients. HA also had fewer complications and lower mortality rates. Therefore, the conclusion is that HA should be considered over PFN for treating intertrochanteric fractures in elderly patients, especially those over 90 years old.
The shoulder joint consists of three bones and three joints. It has a ball and socket configuration that allows for movement in multiple axes. The joint is supported by strong ligaments and muscles like the rotator cuff. Common conditions affecting the shoulder include tendonitis, bursitis, and instability from injuries like dislocations. Pain arises from damage to joint structures innervated by nerves like the axillary nerve.
This document summarizes the laboratory diagnosis of hepatitis B. It discusses the assessment and staging of chronic HBV infection through serological markers, viral load measurement, and liver disease severity evaluation. Rapid diagnostic tests, ELISA/EIA, CLIA, and nucleic acid testing are described for diagnosing HBV. Treatment options including tenofovir and entecavir are outlined. Monitoring during and after treatment includes assessing disease progression, side effects, and hepatocellular carcinoma. Prevention of mother-to-child transmission through vaccination and special considerations for pregnancy and other comorbidities are also covered.
This document provides an overview of emerging viral infections, including their classification, factors contributing to their emergence, prevention, and latest updates. It discusses several important emerging and re-emerging viruses such as influenza virus, dengue virus, Nipah virus, Rift Valley fever virus, monkeypox virus, and more. For each virus, it covers topics like clinical presentation, diagnosis, treatment, vaccination, and geographic distribution. It emphasizes that many emerging viruses are RNA viruses that can mutate and evolve rapidly, facilitating their spread and adaptation to new environments and hosts.
1. The document discusses various culture media used for cultivating microorganisms from clinical specimens. It describes the composition and purpose of different types of media including basal media, enriched media, selective media, and transport media.
2. Specific media are described for cultivating gram positive cocci like Staphylococci and Streptococci. Mannitol salt agar, tellurite glycine agar, and DNase test agar are discussed for isolating and identifying Staphylococcus. Todd Hewitt broth and crystal violet blood agar are mentioned for Streptococci.
3. Modified Thayer Martin medium and modified New York City medium are highlighted as selective media used for isolating Neisseria
The document discusses the human microbiome and its relationship to human health. It defines the microbiome as the collection of microorganisms that live on and inside the human body. The microbiome plays important roles in metabolism, immune function, and protecting against pathogenic bacteria. Imbalances or dysbiosis in the normal microbiota have been linked to various diseases. The human microbiome project aims to better characterize human microbiota and understand its role in health and disease.
This document provides information on different types of rashes that can accompany febrile illness. It begins with an introduction on febrile rashes and their diagnostic approach. It then describes various rash patterns such as maculopapular, vesicular, and urticarial rashes. Specific infectious diseases that can present with rashes like measles, rubella, and scarlet fever are discussed. It concludes by summarizing diagnostic tests and distinguishing features of different febrile rashes.
Vancomycin-resistant enterococci (VRE) are a significant cause of hospital-acquired infections. Enterococcus faecium and Enterococcus faecalis are the most common clinical species. VRE have acquired resistance to vancomycin through the alteration of peptidoglycan precursors in cell wall synthesis. The vanA and vanB genotypes are most prevalent and confer high-level resistance. Treatment involves combination therapy with cell wall active agents and aminoglycosides. Early detection of VRE enables timely infection control measures to limit spread.
Travel vaccination in Manchester offers comprehensive immunization services for individuals planning international trips. Expert healthcare providers administer vaccines tailored to your destination, ensuring you stay protected against various diseases. Conveniently located clinics and flexible appointment options make it easy to get the necessary shots before your journey. Stay healthy and travel with confidence by getting vaccinated in Manchester. Visit us: www.nxhealthcare.co.uk
Osteoporosis - Definition , Evaluation and Management .pdfJim Jacob Roy
Osteoporosis is an increasing cause of morbidity among the elderly.
In this document , a brief outline of osteoporosis is given , including the risk factors of osteoporosis fractures , the indications for testing bone mineral density and the management of osteoporosis
Integrating Ayurveda into Parkinson’s Management: A Holistic ApproachAyurveda ForAll
Explore the benefits of combining Ayurveda with conventional Parkinson's treatments. Learn how a holistic approach can manage symptoms, enhance well-being, and balance body energies. Discover the steps to safely integrate Ayurvedic practices into your Parkinson’s care plan, including expert guidance on diet, herbal remedies, and lifestyle modifications.
These lecture slides, by Dr Sidra Arshad, offer a simplified look into the mechanisms involved in the regulation of respiration:
Learning objectives:
1. Describe the organisation of respiratory center
2. Describe the nervous control of inspiration and respiratory rhythm
3. Describe the functions of the dorsal and respiratory groups of neurons
4. Describe the influences of the Pneumotaxic and Apneustic centers
5. Explain the role of Hering-Breur inflation reflex in regulation of inspiration
6. Explain the role of central chemoreceptors in regulation of respiration
7. Explain the role of peripheral chemoreceptors in regulation of respiration
8. Explain the regulation of respiration during exercise
9. Integrate the respiratory regulatory mechanisms
10. Describe the Cheyne-Stokes breathing
Study Resources:
1. Chapter 42, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 36, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 13, Human Physiology by Lauralee Sherwood, 9th edition
One health condition that is becoming more common day by day is diabetes.
According to research conducted by the National Family Health Survey of India, diabetic cases show a projection which might increase to 10.4% by 2030.
Adhd Medication Shortage Uk - trinexpharmacy.comreignlana06
The UK is currently facing a Adhd Medication Shortage Uk, which has left many patients and their families grappling with uncertainty and frustration. ADHD, or Attention Deficit Hyperactivity Disorder, is a chronic condition that requires consistent medication to manage effectively. This shortage has highlighted the critical role these medications play in the daily lives of those affected by ADHD. Contact : +1 (747) 209 – 3649 E-mail : sales@trinexpharmacy.com
share - Lions, tigers, AI and health misinformation, oh my!.pptxTina Purnat
• Pitfalls and pivots needed to use AI effectively in public health
• Evidence-based strategies to address health misinformation effectively
• Building trust with communities online and offline
• Equipping health professionals to address questions, concerns and health misinformation
• Assessing risk and mitigating harm from adverse health narratives in communities, health workforce and health system
Muktapishti is a traditional Ayurvedic preparation made from Shoditha Mukta (Purified Pearl), is believed to help regulate thyroid function and reduce symptoms of hyperthyroidism due to its cooling and balancing properties. Clinical evidence on its efficacy remains limited, necessitating further research to validate its therapeutic benefits.
Our backs are like superheroes, holding us up and helping us move around. But sometimes, even superheroes can get hurt. That’s where slip discs come in.
6. VEEBOT
• Portable robotic device for
automated phlebotomy
• near infra-red visualisation
producing stereoimages and
robotic kinematics introducing
needle and effecting
venepuncture.
• Accuracy,safety,precision
• able to locate small peripheral
vein
9. PTTS....
ADVANTAGES
• FAST
• EASY
• LESS LABOUR
DISADVANTAGES
• HEMOLYSIS OF BLOOD
SAMPLES AND CAN
INTERFERE WITH RESULTS
• COST OF INSTALLATION
AND MAINTENANCE
10. AUTOMATION IN GRAM STAINING
• Smears are prepared and introduced into enclosed system
• 1) specimen fixation
2) stained with crystal violet and thenwashed by deionized water
3) stained with iodine and then washed by deionized water
4) decolorized and counterstained simultaneously by acetone safranin
solution
5) dried by the system.
• Eg :: 1)PREVI color automated gram stainer 2)Aerospray gram
series slide stainer
11. differences....
• Fixation 1]heat
2] alcohol 95%
• Staining methods
1]Bath stainers-chance of carry over of stain
2]Spray stainers-less wastage
3]cuvette designed staining
• Capacity- 30-120/hour
• Turn over time-3-5 min
12.
13. AUTOMATED URINE ANALYZER
• UF-100 AND UF-50
• IQ 200 AUTOMATED URINE MICROSCOPY ANALYZER
• IRIS FLOW VIDEOMICROSCOPY
14. UF-100 ,UF- 50,uf 1000i
• Laser based flow cytometry along with impedance detection ,forward
light scatter and fluorescence to identify cells
• two dyes are used
1] phenathridine - DNA
2]carbocyanine - -ve charged cell membrane
nuclear membrane
mitochondria
15. • The system aspirates 0.8 mL of urine .
• analyse cells [erythrocyte,leukocytes (WBC) and epithelial cells],
Bacteria and casts .
• use electrical impedance for volume
• forward light scatter for size
• uses a couple of fluorescent dyes for nuclear and cytoplasmic
characteristics
• The formed elements are categorised in a two-dimensional space
(scattergrams) on the basis of their size, shape, volume,and staining
characteristics.
16. IQ 200 AUTOMATED URINE MICROSCOPY
ANALYZER
• The sample is mixed,aspirated to a flow cell where 500
photomicroscopic images are taken
• Auto particle recognition system analyses the cells in the
photomicrograph based on cell size,shape ,texture and contrast
17.
18.
19. AUTOMATION IN SPECIMEN PROCESSING
• The currently available speciemen processors include:
1]Innova processor -BD diagnostics
2]InoqulA fullautomation/manual automation [FA/MI] specimen
processing device - BD Diagnostics
3]Previ Isola Automatic plate streaker-Biomerieux
4] Walk Away Specimen Processor[WASP]-Cogan diagnostics
• Process LIQUID -BASED SPECIMENS
20. INNOVA PROCESSOR
• 5 Drawer * 40 tubes = 200 samples
• 270 agar plates
• Library of streaking patterns
• 1,10,30 microlitre nichrome loop
• specimen can be added as they arrive in lab
• automatic decapper
21. InoqulA FA/MI
• FOR
Slide preparation
autoinoculation of liquid specimen
manual inoculation of other specimens
• inoculation using magnetic beads
• different atmospheres of incubation
• more isolated colonies than manual
22.
23. PREVI ISOLAAUTOMATED PLATE STREAKER
• Uncapped bottles
• disposable applicator and pippette for each plate and specimen
• single streak pattern-radial
• 180 plates/hr
• streaked plates are ejected to stacks
• more isolated colonies
24.
25. WASP
• SCARA Robot to move specimens and plates
• Gram smear preparatory module
• 320-370 plates
• automatic loop changer -1,10,30 microL loops
• full library of streaking pattern
• streaked plates into stacks
• half plates can be streaked and labelled.
29. BActerial Rapid Detection using Optical scatter
Technology (BARDOT)
• irradiation of bacterial colonies grown in a Petri dish with a
red laser to generate light scattering patterns. The light
scattering patterns are dependent on the three-
dimensional (3D) morphology of bacterial colonies.
• distinguished Listeria, Staphylococcus, Salmonella,
Vibrio, and Escherichia with classification accuracy of 90–
99% . Five species of Listeria , three species of Vibrio [7],
and seven serogroups of E. coli have been discriminated
with the accuracy of >91%, >96%, and >81%,
respectively.
30.
31. IDENTIFICATION SYSTEMS
• API 20E/NE identification system
• BBL CRYSTAL
• MicroScan Walkaway
• Vitek system
• Sensititre Gram negative auto identification system
• Phoenix system
• Omnilog ID system
32. API (ANALYTICAL PROFILE INDEX)
IDENTIFICATION SYSTEM
• Packaged system for identification of
-Enterobacteriaceae -20 E ID
-non fermentative bacilli -20 E
20 NE system
-Gram positive cocci-API Staph Ident
-API Staph
-API ID 32 Staph
33. API 20 E SYSTEM
• for identification of Enterobacteracea
and non fermenters like
P.aeruginosa,S.maltophila,Acinetobact
er species
• Consists of a plastic strip with 20
cupules containing dehydrated
substrates and a plastic incubation
chamber
• Same day identification-5 hrs or after
24-48 hrs( more accuracy and
precision).
36. BBL CRYSTAL ENTERIC /ON FERMENTER ID
SYSTEM //GRAM POSITIVE IDS
• MINIATURISED
IDENTIFICATION
SYSTEM
• consists of fluorogenic and
chromogenic substrates
• reproducibility 96.3%-
100%
• accuracy of 96.9%
37. VITEK SYSTEM
• VITEK LEGACY SYSTEM AND VITEK 2 SYSTEM
• The VITEK 2 is an automated microbiology system utilizing growth-
based technology.
• 3 formats -VITEK 2 compact-industrial microbiology
-VITEK 2
- VITEK 2 XL
38. VITEK 2 and VITEK 2 XL
more focused on the clinical microbiology laboratory
provide increased levels of automation and capacity for higher volume
laboratories.
They also provide an option of automatic pipetting and dilution for
antimicrobial susceptibility testing
39. COMPONENTS
• Integrated modular system
1] filling - sealer unit
2]reader-incubater
3]computer control module
4]data terminal
5]multicopy printer
40. Reagent Cards
• 64 wells - each contain an individual
test substrate. Substrates measure
various metabolic activities such as
acidification, alkalinization, enzyme
hydrolysis, and growth in the presence
of inhibitory substances
41. TYPES OF REAGENT CARDS
• 1. GN - Gram-negative fermenting and non-fermenting bacilli
• 2. GP - Gram-positive cocci and non-spore-forming bacilli
• 3. YST - yeasts and yeast-like organisms
• 4. BCL - Gram-positive spore-forming bacilli
42. • Suspension Preparation
A sterile swab or applicator stick is used to transfer a sufficient number
of colonies of a pure culture and suspend the microorganism in 3.0 mL
of sterile saline in a plastic test tube.
The turbidity is adjusted accordingly and measured using a turbidity
meter called the DensiChekTM
44. • A test tube containing the
microorganism suspension is placed
into a special rack (cassette) and the
identification card is placed in the
neighboring slot .
• The cassette can accommodate up to
10 tests (VITEK 2 Compact) or up to
15 tests (VITEK 2 and VITEK 2 XL).
45. • The filled cassette is placed either
manually (VITEK 2 compact) or
transported automatically (VITEK 2
and VITEK 2 XL) into a vacuum
chamber station. After the vacuum is
applied and air is re-introduced into
the station, the organism suspension
is forced through the transfer tube
into micro-channels that fill all the
test wells
46. Card Sealing and Incubation
• Inoculated cards are passed by a mechanism, which cuts off the
transfer tube and seals the card prior to loading into the carousel
incubator. The carousel incubator can accommodate up to 30 or up to 60
cards. All card types are incubated on-line at 35.5 + 1.0ºC.
• Each card is removed from the carousel incubator once every 15
minutes, transported to the optical system for reaction readings, and then
returned to the incubator until the next read time.
47. Optical System
A transmittance optical system allows interpretation of test reactions
using different wavelengths in the visible spectrum. During incubation,
each test reaction is read every 15 minutes to measure either turbidity or
colored products of substrate metabolism. Results are interpreted at 3 hr
of incubation.
48. MALDI-TOF
• MATRIX ASSISTED LASER DESORPTION/IONISATION -TIME OF
FLIGHT MASS SPECTROMETRY
• Identification Based on PEPTIDE MASS FINGERPRINT
• commercially available systems include
-Vitek MS -[bioMerieux]
-MALDI Biotyper-[Bruker]
49. • SAMPLES
Culture -bateria,mycobacteria,yeasts,molds
sediment from positive blood culture
sediment from specimens like urine,CSF, stool
• MATRIX SOLUTION USED
alpha cyano 4 hydroxy cinnamic acid
sinapinic acid(3,5-dimethoxy-4hydroxycinnamic acid)
2,5 dihydroxy benzoic acid
• SOLVENT USED
acetonitrile
trifluoroacetic acid
ethanol
highly purified water
51. • Ionised particles to mass
spectrometer vaccum chamber
• migration due to potential
difference
• and velocity of migration depends
on the mass to charge ratio
52. • TOF- determined by arrival of different molecules at the
detector
• Summation of TOF produce a spectrum
53. DISADVANTAGES
• High capital cost
• Limitations:
Shigella × E.coli
Streptococcus mitis
ADVANTAGES
• Less labor
• less time to identification
• less materials used
• ※good clinical impact
54.
55. RESULT INTERPRETATION
• Score value
≥2.3 - reliable genus and species
ID
2.0-2.29 - genus reliable species
probable
1.7-1.99 - genus probable
≤1.7 - not reliable
56. MALDI TOF IN BACTERIOLOGY
• Identification of bacteria from clinical specimens,culture plates,
• bacterial strain typing and taxonomy
• detect antimicrobial resistance-beta lactamase production,carbapenemase
activity.detect the antibiotic mass alteration due to chemical modification
• detection ,identification and inactivation of biological warfare
• food and water safety
57. APPLICATIONS IN VIROLOGY
• Diagnosis of Influenza,entero virus,HPV,herpes,hepatitis viruses
• genotyping of hepatitis B $ C virus,JC polyoma virus
• for typing ,subtyping,& tracing the lineage of human influenza virus
• detection of mutation in hepatitis B virus
• drug resistance of gancyclovir in CMV
58. APPLICATION IN MYCOLOGY
• Less advanced - drug resistance and fungal strain typing is not advanced
• Fungi identified are Candida,Cryptococcus,
Dermatophytes,Fusarium,Aspergillus,Pencillium.
59. SENSITITRE AUTO IDENTIFICATION
SYSTEM
• The sensititre automated reading and incubation
system(ARIS)(TREK diagnostic system) is an automated
system
• uses fluorescent technology to detect bacterial growth
and enzyme activity.
• Consists of 32 biochemical tests and fluorescent tests.
60. • Each biochemical test
medium along wt appropriate
fluorescent indicator is dried
into individual wells of
sensititre plate.
• Tests are read for the
presence or absence of
fluorescence.
• Results are transmitted to
computer for analysis and
identification.
61. Results –read after 5 hr
incubation
If identification cannot be
obtained---read after
overnight reincubation.
62. THE PHOENIX SYSTEM
• A newly developed fully automated identification and
antimicrobial susceptibility test system.
• comprised of disposable panels that combine
identification and AST .
• perform automatic reading at every20 mts,during
incubation.
63. • Gram negative identification segment uses 45
biochemical substrate
• 16 fluorogenic + 14 fermentation + 8 carbon source + 5
chromogenic + urea and ornithine.
• Identify aerobic GNB in 2-12 hrs.
• Instrument monitor both fluorescent level and visible
spectral changes ,interprets the result.
64. THE OMNILOG ID SYSTEM
• Fully automated system.
• Uses carbon sourse utilization method.
• Simultaneously incubates ,read and interprets the
microplates.
• Continuously process the sample but allows complete
access at any time during sample run.
• Allows organism to be incubated at their optimal
temperature
65.
66. • Reads microplate after 4 hr--Pattern is compared to the
identification data base--and an ID is called if enough
positive reactions have developed.
• If no result is obtained after 6 hr,instrument automatically
continues to incubate the plates and read after 16 hr-24
hr.
• database contain 1400 different organisms ,including
501 gram negative species.
67. VIDAS AND MINIVIDAS
PRINCIPLE
• The assay principle combines an enzyme immunoassay
competition method with a final fluorescent detection
• The Solid Phase Receptacle (SPR®) serves as the solid
phase as well as the pipetting device for the assay.
• Reagents for the assay are ready-to-use and predispensed
in the sealed reagent strips.
• All of the assay steps are performed automatically by the
instrument. The reaction medium is cycled in and out of
the SPR several times.
68. • The sample is collected and transferred into the well
containing an alkaline phosphatase-labeled antibody (conjugate).
• The antigen present in the sample
and the antigen coated on the interior of the SPR
compete for the available sites on the specific antibody conjugated
to alkaline phosphatase.
During the final detection step, the substrate
(4-Methyl-umbelliferyl phosphate) is cycled in and out of the SPR.
69. The conjugate enzyme catalyzes the hydrolysis
of this substrate into a fluorescent product
(4-Methyl-umbelliferone), the fluorescence of which is
measured at 450 nm. The intensity of the fluorescence is
inversely proportional to the concentration of antigen
present in the sample
70. SHERLOCK MICROBIOLOGICAL IDS
• gas chromatography (GC) system dedicated to bacteria
identification by fatty acid methyl ester (FAME) analysis is
the Sherlock Microbial Identification System (MIS)
• The principle of the FAME method rests upon the
assumption that some microorganisms have typical
cellular fatty acid compositions, which can be compared
with the mean fatty acid composition of the strains used to
create the library. After comparison, the identities of
unknown microorganisms are determined.
71.
72. NOVEL TECHNOLOGIES IN ID
• Surface Plasmon Resonance imaging (SPRi) is an optical
detection technique enabling the real-time and label-free
monitoring of molecular interactions occurring on metallic
layers. Recently, SPRi has been coupled to protein
microarrays to detect food-related bacterial pathogens in
small (about 1 mL) sample volumes. The successful
detection of live bacteria was possible by choosing
antibodies specific to antigens expressed by the bacteria
and located on the cell surface
73. Live bacteria are
captured on
microarrayed specific
antibodies (spotted in
triplicate onto the
biochip surface) during
the enrichment step.
SPRi data are treated
and plotted as variations
of light reflectivity (ΔR
(%)) over time for each
region-of-interest
(corresponding to
antibody spots arrayed
on the sensor).
75. DIRECT RAPID ANTIMICROBIAL SUSCEPTIBILITY
TESTING[DRAST]
• AST from positive blood culture bottles
• microscopic image analysis
• Growth detection and time lapse for MIC calculation
76.
77. MAC CHIP
• Micro Fluidic Agarose Channel
chip
• Rapid AST by single cell tracking
78. Total Lab Automation (TLA)
3 TLA system
1) Kiestra TLA
2) FMLA (Full Microbiology Laboratory Automation)
3) WASP lab
80. KIESTRA TLA
5 modules-linked by conveyer
Sorter A
Barcod A
Inoql A –processor
Read A –incubator with digital imaging equipment
ergonomics A –work benches
85. WASP
CO2 & nonCO2 incubator
Linked by conveyer system
Image acquisition station-capture image
Plate with growth ---reloaded on WASP lab for automated
broth inoculation and Kirby bauer disk dispensing
86.
87. AUTOMATION IN BLOOD CULTURE
CULTURE METHODS
RAPID
IDENTIFICATION
METHODS
OLD NEW
[CONT. MONITORING]
OLDER METHODS
BACTEC BACTEC 9240/9120
NOVEL METHODS
BACT/ALERT
1]Lysis centrifugation intrinsic
fluorescence
2]Integrated Comprehensive
direct droplet
TREK ESP
88. BACTEC
• Early automated method
• Radiometric method
• non-continuous monitoring
• 3-5m1 sample is incubated at 35.37°C in a sealed
rubber septum vial with a liquid14 C-labelled sterilized
substrate with an activity of: 2 uCi per vial
• If bacteria are present they metabolize carbohydrate or
protein, the components of the substrate as energy
source, releasing14 co2 by catabolizing glucose or by
decarboxylation of aminoacid produced during incubation.
89. • 14 C02 produced during the incubation period is then aspirated from the
test vial through sterilizing filter into the ionization chamber, the
electrometer present in Bactec unit then measures the current produced
in the ionization chamber.
• Growth index (GI) : The amount of 14 CO2 liberated is proportional to
the amount of the bacterial growth in the nutrient media. A reading of
100 GI corresponds to 0.025 microcurie of14 C.
• A threshold GI may be set which is usually 30 for aerobic vials and 20
for anaerobic vials, a reading above threshold level indicates the
presence of bacteria.
90. BACTEC 9000 series-
• continuous monitoring system
• CO2 production is detected
• signals when growth detection
• tube with growth can be remov manually and can be
proceeded
91. CONSTITUENTS IN BACTEC VIAL
• Different media for aerobic culture,anaerobic
culture,fungal culture and paediatric sample
• anticoagulant - 0.025 -0.05% sodium polyanethol
sulfonate-toxic to certain bacteria eg: Neisseria
• Resins-Neutralise antimicrobial activity
except:imipenem,cilastatin
• CO2 & O2 (N2 in Anaerobic medium )
• sensor for detection of fluorescence
92. The BacT/Alert blood culture system
First continuous monitoring blood
culture system
Each blood culture bottle -10 ml blood
Data unit- A cabinet about the size of a
small refrigerator. Serves as self
contained incubator,shaker and
detection device. Hold 240 or 120
bottles.
Bottle is placed bottom down in to the
receiving well in the data
unit,directed by a bar code on bottle
label.
93. Bottom of bottle has a colour sensor --separated from blood
broth mixture by a unidirectional CO2 permeable
membrane.
Light sensitive detector is also present .
94. Micro organism grow in blood broth mixture
CO2 liberated
CO2 + H20 → H2C03↔ [H+] + {HCO3-}
{ACIDIC pH]
Colour of sensor turns from green to yellow
95. • At 10 mt intervals , light beam from emitting
diode (1 for each well) is projected through an
excitation filter to reflect off CO2 sensitive
sensor.Reflecting light is directed through an
emission filter to a photo sensitive detector
• Sufficient CO2----alter the sensor------
visible/audible alert.
• Positive bottle removed for processing.
• 144 times / bottle / day
• data points are plotted as reflectance unis
versus time and result in a growth curve
96. The TREK ESP culture system II
Differs from other 2 in the following ways
1) Production of CO2 is measured manometrically.
2) both gas production and consumption are monitored.
3) change in concentration of H2 and O2 addition to CO2
are detected
101. BACTEC AFB SYSTEM –BACTEC 460
Principle:
Growth medium for culturing Mycobacteria is
supplemented with a substrate labeled with radioactive
carbon (14C).
When Mycobacteria grow in this medium, they
metabolize the labeled substrate and release 14CO2 into
the atmosphere above the medium inside the sealed
bottle.
102. The Bactec 460 instrument tests for the presence 14CO2
by removing the gas from the bottle and transferring it to
an electrometer, where the amount of ionization is
converted to a number called a Growth Index (GI).
The rate and amount of 14CO2 (GI) produced is directly
proportional to the rate and amount of bacterial growth in
the medium
103. PANTA mix to suppress contaminant growth
C14 labeled palmitic acid-growth detector
Vial is inoculated with 0.5 ml processed specimen and
incubate at 35⁰C.
When designated period of incubation is over,3 days
usually,vials are placed on track of BACTEC 460
instrument in preparation for reading
104. A growth index more than 10 ---positive
Detection time of M.tuberculosis—9-14 days
Rapid growers-less than 7 days
Disadvantage:
High cost
inability to observe colony morphology
Over growth by contaminants
Need for disposal of radioactive material
105. Can perform rapid drug susceptibility study-adv
Can differentiate M.tuberculosis and bovis from
nontuberculous mycobacteria using blood culture vial
containing NAP(P-nitro-alpha acetylamino-beta hydroxy
propriophenone)
M.tb and bovis can not grow in NAP medium.
106. MYCOBACTERIA GROWTH INDICATOR TUBE (MGIT) &
MGIT 960
MGIT system consists -16 x 100 mm round bottomed glass
tubes –4 ml modified 7H9 broth base + 0.5 ml OADC
enrichment(oleic acid ,bovine albumin,dextrose and
catalase)+ 0.1 ml PANTA antibiotic mixture.
A fluorescent compound is embedded in silicone on the
bottom of the tube---sensitive to dissolved O2 in broth.
107. Presence of O2 in uninoculated medium serves to quench
fluorescence.
As bacteria grows ----consumes O2---fluorescence is
unmasked----detected uv light(woods lamp)
Growth may be also detected by non homogenous turbidity
or small grains in culture medium.
108. 0.5 ml of specimen is added to broth—incubate-tubes are
read every other day starting on 2 nd day after
inoculation—read with woods lamp,placing the tube b/w a
positive(Na sulphite solution) and negative control(un
inoculated medium)
Positive tubes—stained for AFB
Negative tubes---reincubated and observed at regular
interval for up to 6 wks.
109. MGIT 960 I --- a non radiometric ,automated system----uses
BD BBL MGIT media and sensor to detect the
fluorescence that is visually interpreted .
Holds 960 plastic tubes –continuously monitored
DST possible
110. MB/Bac T Mycobacteria detection system
Similar to BacT/Alert blood culture system.
Bottle—10 ml enhanced middlebrook 7H9broth
CO2,N2,O2 under vaccum.
MAS—0.5 ml+ 0.5 ml specimen
Bottom sensor changes from dark green to yellow in
presence of CO2.
Reflected light is used to monitor CO2 production.
DST can be done
111. The ESP culture system II
Adaption of ESP blood culture system
Each bottle when placed in a special drawer in incubation
module,is attached to a senor.
Each bottle is continuously monitored for any change in gas
pressure due to metabolic activity of microorganisms in
broth.
Significant changes may be signalled early ,from the
consumption of O2,or later with production of gas by
metabolism
112. The bottle contain modified middlebrook 7H9
medium,casitone ,glycerol and cellulose sponges.
Sponges—growth platform for mycobacteria like alveoli.
Prior to inoculation add antibiotic mixture –PVNA—
polymyxin B ,vancomycin,nalidixic acid,amphotericin B.
DST possible
113. The BACTEC MYCO/F LYTIC
Bottle contain a lytic agent to release mycobacteria that are
phagocytosed by WBC.
Incubated and monitored automatically and continuously
like other BACTEC blood culture bottle.
Good culture media for bacteria and fungi in bloodstream.
114. BacT/Alert 3D
Recovery of a wide range of pathological organisms,
including bacteria, yeasts and mycobacteria.
The BacT/ALERT 3D is the most compact, modular and
flexible blood culture system available, providing a single
platform for the recovery of microorganisms from blood,
sterile body fluids and mycobacterial specimens, both
respiratory and non-respiratory.