A Fully Automated POCKIT Central PCR System for Evaluation of the Infectious ...Simon Chung - genereach
This document discusses using PCR to evaluate the uniformity of infectious bronchitis vaccination in chickens. It finds:
1) A POCKIT Central PCR system can detect the IB vaccine virus in tracheal swabs up to 4 weeks after spraying vaccination at hatcheries.
2) Sequence analysis showed nucleic acids detected by the PCR system matched the vaccine used in the study.
3) A PCR positive rate of 40% or higher on day 7 after vaccination could be used as a cutoff for evaluating uniformity, with regular monitoring needed below that threshold.
The study demonstrates PCR is an effective quality control tool for checking IB vaccination uniformity. The portable POCKIT system makes this feasible for poultry grow
A field-deployable RT-PCR system performs equivalently to real-time RT-PCR in...Simon Chung - genereach
A field-deployable RT-PCR system was found to perform equivalently to real-time RT-PCR in detecting type 2 porcine reproductive and respiratory syndrome virus (PRRSV). The field-deployable system provided results within 2 hours compared to 2.5 days for laboratory RT-PCR. Testing 50 vaccinated and 50 unvaccinated piglets over 11 weeks showed 96.25% agreement between the two methods. The field-deployable PCR system has potential for timely PRRSV detection and biosecurity management at points of need.
A Fully Automated Sample-to-result PCR System for Detecting Infectious DiseasesSimon Chung - genereach
This document describes GeneReach Biotechnology Corporation's POCKIT Central system, a fully automated sample-to-result PCR system for infectious disease detection. The POCKIT Central can detect multiple pathogens from a single sample in less than 2 hours, including dengue virus serotypes. It has been validated against qPCR with equivalent sensitivity and specificity for dengue virus detection and subtyping. The portable and easy-to-use POCKIT Central provides a rapid and accurate molecular diagnostic solution for point-of-care infectious disease testing.
The document discusses the application of polymerase chain reaction (PCR) in detecting infectious diseases. Specifically, it discusses:
1) PCR is a widely used nucleic acid amplification technique that can replicate a specific DNA region millions of times, allowing detection of small amounts of DNA or RNA from infectious pathogens.
2) PCR has revolutionized clinical infectious disease diagnosis by enabling rapid and accurate detection of viruses, bacteria, and other pathogens from patient samples.
3) The document provides examples of how PCR has been used to detect several infectious diseases, including hepatitis B, hepatitis C, human papillomavirus, HIV, influenza, and the novel coronavirus COVID-19.
SARS-CoV-2 (COVID-19) is the virus responsible for respiratory disease caused by a novel (new) coronavirus that was first detected in Wuhan City, Hubei Province, China, later causing a global pandemic.
The virus is contagious and can be spread from humans to humans primarily through the exchange of mucus droplets that are expelled through sneezes or coughs.
The SARS-CoV-2 is a severe acute respiratory syndrome coronavirus. This outbreak is an important reminder that the global community must strengthen national and international programs for detection and response to future disease outbreaks.
Corona is here to stay and it is predicted that over 70% of population will get the infection (fortunately not all will fall sick or very sick). (Recovery rate of over 74% & Death rate around 2%).
A lot of confusion exists regarding testing for covid and what test to do, when and how to interpret these tests.
Compiled by Dr. Narendra Malhotra
RT PCR is too slow for effective control of spread of cov 2 infection, rapid antigen test by giving results in less than 30 minutes can help identify infected persons leading to quick isolation.Lack of sensitivity can be compensated by repeating RAT after a day or so.
This is a presentation giving an overview of the GeneXpert DX system for detection of MTB. The assay described in this presentation is the MTB/RIF test.
A Fully Automated POCKIT Central PCR System for Evaluation of the Infectious ...Simon Chung - genereach
This document discusses using PCR to evaluate the uniformity of infectious bronchitis vaccination in chickens. It finds:
1) A POCKIT Central PCR system can detect the IB vaccine virus in tracheal swabs up to 4 weeks after spraying vaccination at hatcheries.
2) Sequence analysis showed nucleic acids detected by the PCR system matched the vaccine used in the study.
3) A PCR positive rate of 40% or higher on day 7 after vaccination could be used as a cutoff for evaluating uniformity, with regular monitoring needed below that threshold.
The study demonstrates PCR is an effective quality control tool for checking IB vaccination uniformity. The portable POCKIT system makes this feasible for poultry grow
A field-deployable RT-PCR system performs equivalently to real-time RT-PCR in...Simon Chung - genereach
A field-deployable RT-PCR system was found to perform equivalently to real-time RT-PCR in detecting type 2 porcine reproductive and respiratory syndrome virus (PRRSV). The field-deployable system provided results within 2 hours compared to 2.5 days for laboratory RT-PCR. Testing 50 vaccinated and 50 unvaccinated piglets over 11 weeks showed 96.25% agreement between the two methods. The field-deployable PCR system has potential for timely PRRSV detection and biosecurity management at points of need.
A Fully Automated Sample-to-result PCR System for Detecting Infectious DiseasesSimon Chung - genereach
This document describes GeneReach Biotechnology Corporation's POCKIT Central system, a fully automated sample-to-result PCR system for infectious disease detection. The POCKIT Central can detect multiple pathogens from a single sample in less than 2 hours, including dengue virus serotypes. It has been validated against qPCR with equivalent sensitivity and specificity for dengue virus detection and subtyping. The portable and easy-to-use POCKIT Central provides a rapid and accurate molecular diagnostic solution for point-of-care infectious disease testing.
The document discusses the application of polymerase chain reaction (PCR) in detecting infectious diseases. Specifically, it discusses:
1) PCR is a widely used nucleic acid amplification technique that can replicate a specific DNA region millions of times, allowing detection of small amounts of DNA or RNA from infectious pathogens.
2) PCR has revolutionized clinical infectious disease diagnosis by enabling rapid and accurate detection of viruses, bacteria, and other pathogens from patient samples.
3) The document provides examples of how PCR has been used to detect several infectious diseases, including hepatitis B, hepatitis C, human papillomavirus, HIV, influenza, and the novel coronavirus COVID-19.
SARS-CoV-2 (COVID-19) is the virus responsible for respiratory disease caused by a novel (new) coronavirus that was first detected in Wuhan City, Hubei Province, China, later causing a global pandemic.
The virus is contagious and can be spread from humans to humans primarily through the exchange of mucus droplets that are expelled through sneezes or coughs.
The SARS-CoV-2 is a severe acute respiratory syndrome coronavirus. This outbreak is an important reminder that the global community must strengthen national and international programs for detection and response to future disease outbreaks.
Corona is here to stay and it is predicted that over 70% of population will get the infection (fortunately not all will fall sick or very sick). (Recovery rate of over 74% & Death rate around 2%).
A lot of confusion exists regarding testing for covid and what test to do, when and how to interpret these tests.
Compiled by Dr. Narendra Malhotra
RT PCR is too slow for effective control of spread of cov 2 infection, rapid antigen test by giving results in less than 30 minutes can help identify infected persons leading to quick isolation.Lack of sensitivity can be compensated by repeating RAT after a day or so.
This is a presentation giving an overview of the GeneXpert DX system for detection of MTB. The assay described in this presentation is the MTB/RIF test.
Real time rt-pcr Test molecolare per covid-19 (Sars-CoV-2) Protocollo UfficialeIULALaboratorioAnali
Protocollo ufficiale per la ricerca e amplificazione di RNA virale mediante tecnica PCR-RealTime per l'individuazione del virus Covid-19
Per maggiori informazioni: https://www.laboratorioiula.com
A Field-Deployable Insulated Isothermal PCR-Based System for Rapid and Sensit...Simon Chung - genereach
POCKIT Central PCR System for Detecting African Swine Fever Virus in Vietnam
The document describes a study evaluating the POCKIT Central PCR system for detecting African Swine Fever Virus (ASFV) in Vietnam. The system provides fully automated sample-to-answer detection of ASFV in 85 minutes using cartridges that integrate nucleic acid extraction and PCR. The study found the POCKIT Central system performed equivalently to the OIE reference real-time PCR method, with high analytical sensitivity and specificity. The automated system reduces hands-on time and human error compared to traditional PCR methods. It provides a simple workflow for rapid on-site detection of ASFV to help control the ongoing spread of the disease
neoplex pcr test kit [genematrix advantages of neo_plex covid-19 Rapid Test K...HK HuZef
NeoPlexTM COVID-19 has advantages over PowerChek 2019-nCoV for PCR-based detection of COVID-19 including higher sensitivity detecting down to 10-5 compared to 10-3 for PowerChek, targeting two genes (RdRp and N) specific to COVID-19 versus one broadly reactive gene, and including an internal process control for nucleic acid extraction and PCR. NeoPlex also offers advantages of being more user-friendly, requiring fewer PCR machines, allowing testing of more samples per kit, and having longer shelf life and reagent stability.
This document describes a kit for detecting the human pathogen Mycobacterium tuberculosis using PCR. It discusses how the kit allows for isolation of DNA from samples using a spin column method. It then can detect M. tuberculosis through amplification of a region of its genome using provided master mixes and controls. The kit is designed for testing 24 samples at a time and provides high sensitivity and specificity for detection of this important pathogen.
RT2 Profiler PCR Arrays: Pathway-focused Gene Expression Profiling with qRT-P...QIAGEN
This paper evaluates the performance of the newest technique for monitoring the expression of a panel of pathway- or disease-specific genes: the RT2 Profiler PCR Array System. The RT2 Profiler PCR Array System combines the quantitative performance of SYBR® Green real-time PCR with the multiple-gene profiling capabilities of a microarray.
The RT2 Profiler PCR Array is a 96- or 384-well plate containing RT2 qPCR Primer Assays for a set of 84 related genes, plus 5 housekeeping genes and 3 controls. The complete system includes an instrument-specific master mix and an optimized first strand synthesis kit. This paper presents experimental data showing that RT2 Profiler PCR Arrays have the sensitivity, reproducibility, and specificity expected from real-time PCR techniques. As a result, this technology brings focused gene expression profiling to any biological laboratory setting with a real-time PCR instrument.
The document describes RT2 Profiler PCR Arrays, which allow for pathway-focused gene expression profiling using real-time PCR. The PCR Arrays contain primer sets for 84 relevant genes, plus controls. They have been shown to have high sensitivity, specificity, and reproducibility. The complete system includes optimized primer assays, master mixes, and a first strand synthesis kit. Researchers can use pre-designed arrays focused on biological pathways or diseases, or customize arrays as needed.
The COVID-19 pandemic, also known as the corona virus pandemic, is an ongoing pandemic of corona virus disease 2019 (COVID‑19) caused by severe acute respiratory syndrome corona virus 2 (SARS‑CoV‑2).
Though several trials for candidate vaccines and potential therapies are underway, there is currently no cure, and in the absence of either proven effective therapy or a vaccine, diagnostic testing becomes a valuable tool. Testing is our window onto the pandemic and how it is spreading. Without data on who is infected by the virus we have no way of understanding the pandemic. Without this data we cannot know which countries are doing well, and which are just underreporting cases and deaths.
At IntellectPeritus, we have collected around 100 diagnostic kits (used in SARS-COV-2 diagnosis) and around 50 patent documents as sample report and have tried to put value added technology categories to generate specific insights quickly using our deliverable “Innovigence Corona”.
If you are interested in detailed or any customized report for SARS-COV-2 diagnostic kits please feel free to write to us at sales@intellectperitus.com
10 Tips to maximize your Real Time PCR Success - Download the Technical NoteQIAGEN
This document provides 10 tips for maximizing success with real-time PCR. The tips include using high-quality nucleic acid templates, determining template concentration and purity, checking storage conditions, using the optimal template amount, determining reaction efficiency for each primer pair, properly storing primers and probes, preventing contamination, thoroughly mixing reaction components, performing necessary control reactions, and double checking cycler settings.
The Xpert MTB/RIF assay uses molecular beacon probes to detect mutations in the rpoB gene associated with rifampin resistance. It contains five probes that bind to the wild-type rpoB sequence and one probe for an internal sample processing control. If one of the five probes does not bind, it indicates a mutation and rifampin resistance. The assay interprets results based on cycle threshold values and differences in values between probes to determine MTB detection level and presence of resistance. Internal controls verify reagent rehydration, tube filling, and detect inhibition to help prevent false negatives.
The document describes a rapid test kit for detecting canine parvovirus antigen in dog feces. Canine parvovirus causes inflammation of the intestines and other symptoms like vomiting and diarrhea. The test uses lateral flow immunoassay to detect parvovirus antigen in a fecal sample in 5-10 minutes. It has a sensitivity of 98.5% and specificity of 98% compared to ELISA tests. A positive result confirms parvovirus infection while a negative result does not rule it out, as virus shedding may be short.
RT2 Profiler PCR Arrays allow researchers to analyze gene expression profiles for focused panels of genes involved in biological pathways or disease states. The arrays provide laboratory-verified gene assays, integrated controls, and free data analysis software to generate a gene expression profile from a sample in less than 3 hours. Popular arrays analyze pathways such as extracellular matrix and adhesion molecules, WNT signaling, and cancer-related genes. The complete workflow begins with sample preparation and ends with data interpretation and publication of results.
Nucleic Acid Quantification from FFPE Samples – Are You Doing it Right?QIAGEN
Formalin-fixation and paraffin-embedding is a standard method for long-term preservation of tissue biopsies and these stored samples are a valuable tool for studying diseases such as cancer, especially when they are histologically and pathologically well characterized, and follow-up clinical data is available. The quality of nucleic acids extracted from FFPE samples is influenced by a number of factors, including how the samples were handled before, during and after fixation and embedding. Moreover, there are several difficulties when purifying nucleic acids from FFPE samples as the chemicals and temperature used during the process can degrade the DNA.
In this webinar, we will discuss the variability in quantity and purity of DNA purified from FFPE material. We will show data from different quantification and quality control methods, demonstrate the impact of inaccurate quantification on downstream results and discuss how to overcome these challenges.
The importance of controls and novel solutions for successful real-time qPCRQIAGEN
The increasing demand for streamlined, monitored and ultrafast qPCR procedures requires high-performance, real-time quantitative RT and PCR chemistries. Particularly, procedures utilizing generic kits for gene expression analysis should include in-process safety measures to avoid variables and control accuracy of procedures and results. This slidedeck presents innovative solutions for one-step and two-step RT-PCR that significantly enhance performance and reliability in qRT-PCR. The new QuantiNova kit family offers a combination of various integrated safety features to remove variables and prevent artifacts. Internal control RNA, removal of genomic DNA, room temperature set-up capability for RT-PCR and a built-in visual pipetting control verify accurate procedures, ensuring reliable gene expression profiling.
This slidedeck explains the principles of the technologies and shows data demonstrating performance in qRT-PCR. Find out how you can verify accurate performance in qRT-PCR and improve your results!
Neil leblanc f7 rapid methods call amsterdam may 2010sva-slu_oie-cc
The document summarizes a strategic meeting discussing the development and use of multiplex assays using Luminex technology. Methods allow the detection of up to 100 unique assays in a single sample, and have been used to detect and subtype pestiviruses, avian influenza viruses, African swine fever virus, and cytokines. The technology provides a sensitive and specific platform for pathogen identification and characterization applicable to diagnostic laboratories.
RT-PCR is a sensitive technique for detecting and quantifying mRNA. It uses reverse transcriptase to synthesize cDNA from an RNA template, which is then used as a template for PCR amplification using DNA polymerase. RT-PCR can be performed as a one-step or two-step process and is commonly used in clinical microbiology labs to detect RNA viruses from specimens. The COVID-19 RT-PCR test analyzes respiratory specimens for SARS-CoV-2 RNA by amplifying small amounts into DNA to accurately diagnose infections.
This document provides information on several kits from Norgen Biotek Corp. for detecting HIV and related pathogens. It summarizes 3 kits that detect HIV: 1) an HIV quantitative RT-PCR kit that detects HIV RNA and has a reported linear range of 102-106 copies/μL, 2) an HIV proviral DNA PCR kit that detects HIV DNA with a linear range of 102-106 copies/μL, and 3) a plasma/serum HIV RT-PCR kit that isolates RNA from plasma/serum and detects HIV with a linear range of 10-8x106 VP/μL. It also summarizes kits for detecting Cryptococcus neoformans DNA and Pneumocystis jirove
Dr. Jeff Baxter - Lab Testing StandardizationJohn Blue
This document summarizes a study on standardizing Trichomonas foetus DNA testing across multiple laboratories. The study aimed to minimize variables that could influence sample or testing quality. It evaluated pooling of positive samples, different sample preparation methods, and real-time PCR protocols across five feeder labs compared to a central study lab. The study found 95.6% agreement between labs and confirmed 175 of 176 positive samples as T. foetus by DNA sequencing. Pooling was found to potentially miss some positives, with 1:5 pooling missing 4% and 1:3 pooling missing 3.5% of positives. The study supports standardizing sample collection, handling, preparation and PCR analysis to increase testing accuracy and consistency.
Quality Control of RNA Samples - For Gene-Expression Results you Can Rely onQIAGEN
By their very nature RNA molecules, especially mRNA and regulator RNA, are labile and can be highly unstable and sensitive to heat, UV and RNase contamination. The quality, relevance and scientific impact of gene expression results directly depends on the ability to extract RNA without losing any fraction of interest, while preserving the integrity of the biological information it carries. RNA quality control is thus critical to ensure high-quality results and for turning these results into actionable insights with confidence.
In this webinar, we will introduce you to the main parameters influencing RNA-based assays and their respective impact on downstream applications, discuss how to monitor them and cover the advantages of automation for quality control along complex workflows.
A panel was developed using the OpenArray platform to profile common respiratory tract pathogens via PCR. Assays were designed to target viral and bacterial sequences with high specificity and strain coverage. The panel demonstrated high specificity when tested against genomic standards. Pre-amplification improved sensitivity by enhancing detection of low copy targets. The panel provides a customizable and high-throughput tool for respiratory infection research.
Participants of the workshop learn the necessary background information and techniques to diagnose Sars-CoV-2 using the mobile diagnostic laboratory. The laboratory is shipped ready to use with all devices, reagents, certificates, and protocols. After one day of preparation together with a local assistant, a five-day course is given where every step is carried out by each participant. Experts accompany the learning process with written teaching materials, video training, virtual live coaching, and short exams to verify the learned content.
Real time rt-pcr Test molecolare per covid-19 (Sars-CoV-2) Protocollo UfficialeIULALaboratorioAnali
Protocollo ufficiale per la ricerca e amplificazione di RNA virale mediante tecnica PCR-RealTime per l'individuazione del virus Covid-19
Per maggiori informazioni: https://www.laboratorioiula.com
A Field-Deployable Insulated Isothermal PCR-Based System for Rapid and Sensit...Simon Chung - genereach
POCKIT Central PCR System for Detecting African Swine Fever Virus in Vietnam
The document describes a study evaluating the POCKIT Central PCR system for detecting African Swine Fever Virus (ASFV) in Vietnam. The system provides fully automated sample-to-answer detection of ASFV in 85 minutes using cartridges that integrate nucleic acid extraction and PCR. The study found the POCKIT Central system performed equivalently to the OIE reference real-time PCR method, with high analytical sensitivity and specificity. The automated system reduces hands-on time and human error compared to traditional PCR methods. It provides a simple workflow for rapid on-site detection of ASFV to help control the ongoing spread of the disease
neoplex pcr test kit [genematrix advantages of neo_plex covid-19 Rapid Test K...HK HuZef
NeoPlexTM COVID-19 has advantages over PowerChek 2019-nCoV for PCR-based detection of COVID-19 including higher sensitivity detecting down to 10-5 compared to 10-3 for PowerChek, targeting two genes (RdRp and N) specific to COVID-19 versus one broadly reactive gene, and including an internal process control for nucleic acid extraction and PCR. NeoPlex also offers advantages of being more user-friendly, requiring fewer PCR machines, allowing testing of more samples per kit, and having longer shelf life and reagent stability.
This document describes a kit for detecting the human pathogen Mycobacterium tuberculosis using PCR. It discusses how the kit allows for isolation of DNA from samples using a spin column method. It then can detect M. tuberculosis through amplification of a region of its genome using provided master mixes and controls. The kit is designed for testing 24 samples at a time and provides high sensitivity and specificity for detection of this important pathogen.
RT2 Profiler PCR Arrays: Pathway-focused Gene Expression Profiling with qRT-P...QIAGEN
This paper evaluates the performance of the newest technique for monitoring the expression of a panel of pathway- or disease-specific genes: the RT2 Profiler PCR Array System. The RT2 Profiler PCR Array System combines the quantitative performance of SYBR® Green real-time PCR with the multiple-gene profiling capabilities of a microarray.
The RT2 Profiler PCR Array is a 96- or 384-well plate containing RT2 qPCR Primer Assays for a set of 84 related genes, plus 5 housekeeping genes and 3 controls. The complete system includes an instrument-specific master mix and an optimized first strand synthesis kit. This paper presents experimental data showing that RT2 Profiler PCR Arrays have the sensitivity, reproducibility, and specificity expected from real-time PCR techniques. As a result, this technology brings focused gene expression profiling to any biological laboratory setting with a real-time PCR instrument.
The document describes RT2 Profiler PCR Arrays, which allow for pathway-focused gene expression profiling using real-time PCR. The PCR Arrays contain primer sets for 84 relevant genes, plus controls. They have been shown to have high sensitivity, specificity, and reproducibility. The complete system includes optimized primer assays, master mixes, and a first strand synthesis kit. Researchers can use pre-designed arrays focused on biological pathways or diseases, or customize arrays as needed.
The COVID-19 pandemic, also known as the corona virus pandemic, is an ongoing pandemic of corona virus disease 2019 (COVID‑19) caused by severe acute respiratory syndrome corona virus 2 (SARS‑CoV‑2).
Though several trials for candidate vaccines and potential therapies are underway, there is currently no cure, and in the absence of either proven effective therapy or a vaccine, diagnostic testing becomes a valuable tool. Testing is our window onto the pandemic and how it is spreading. Without data on who is infected by the virus we have no way of understanding the pandemic. Without this data we cannot know which countries are doing well, and which are just underreporting cases and deaths.
At IntellectPeritus, we have collected around 100 diagnostic kits (used in SARS-COV-2 diagnosis) and around 50 patent documents as sample report and have tried to put value added technology categories to generate specific insights quickly using our deliverable “Innovigence Corona”.
If you are interested in detailed or any customized report for SARS-COV-2 diagnostic kits please feel free to write to us at sales@intellectperitus.com
10 Tips to maximize your Real Time PCR Success - Download the Technical NoteQIAGEN
This document provides 10 tips for maximizing success with real-time PCR. The tips include using high-quality nucleic acid templates, determining template concentration and purity, checking storage conditions, using the optimal template amount, determining reaction efficiency for each primer pair, properly storing primers and probes, preventing contamination, thoroughly mixing reaction components, performing necessary control reactions, and double checking cycler settings.
The Xpert MTB/RIF assay uses molecular beacon probes to detect mutations in the rpoB gene associated with rifampin resistance. It contains five probes that bind to the wild-type rpoB sequence and one probe for an internal sample processing control. If one of the five probes does not bind, it indicates a mutation and rifampin resistance. The assay interprets results based on cycle threshold values and differences in values between probes to determine MTB detection level and presence of resistance. Internal controls verify reagent rehydration, tube filling, and detect inhibition to help prevent false negatives.
The document describes a rapid test kit for detecting canine parvovirus antigen in dog feces. Canine parvovirus causes inflammation of the intestines and other symptoms like vomiting and diarrhea. The test uses lateral flow immunoassay to detect parvovirus antigen in a fecal sample in 5-10 minutes. It has a sensitivity of 98.5% and specificity of 98% compared to ELISA tests. A positive result confirms parvovirus infection while a negative result does not rule it out, as virus shedding may be short.
RT2 Profiler PCR Arrays allow researchers to analyze gene expression profiles for focused panels of genes involved in biological pathways or disease states. The arrays provide laboratory-verified gene assays, integrated controls, and free data analysis software to generate a gene expression profile from a sample in less than 3 hours. Popular arrays analyze pathways such as extracellular matrix and adhesion molecules, WNT signaling, and cancer-related genes. The complete workflow begins with sample preparation and ends with data interpretation and publication of results.
Nucleic Acid Quantification from FFPE Samples – Are You Doing it Right?QIAGEN
Formalin-fixation and paraffin-embedding is a standard method for long-term preservation of tissue biopsies and these stored samples are a valuable tool for studying diseases such as cancer, especially when they are histologically and pathologically well characterized, and follow-up clinical data is available. The quality of nucleic acids extracted from FFPE samples is influenced by a number of factors, including how the samples were handled before, during and after fixation and embedding. Moreover, there are several difficulties when purifying nucleic acids from FFPE samples as the chemicals and temperature used during the process can degrade the DNA.
In this webinar, we will discuss the variability in quantity and purity of DNA purified from FFPE material. We will show data from different quantification and quality control methods, demonstrate the impact of inaccurate quantification on downstream results and discuss how to overcome these challenges.
The importance of controls and novel solutions for successful real-time qPCRQIAGEN
The increasing demand for streamlined, monitored and ultrafast qPCR procedures requires high-performance, real-time quantitative RT and PCR chemistries. Particularly, procedures utilizing generic kits for gene expression analysis should include in-process safety measures to avoid variables and control accuracy of procedures and results. This slidedeck presents innovative solutions for one-step and two-step RT-PCR that significantly enhance performance and reliability in qRT-PCR. The new QuantiNova kit family offers a combination of various integrated safety features to remove variables and prevent artifacts. Internal control RNA, removal of genomic DNA, room temperature set-up capability for RT-PCR and a built-in visual pipetting control verify accurate procedures, ensuring reliable gene expression profiling.
This slidedeck explains the principles of the technologies and shows data demonstrating performance in qRT-PCR. Find out how you can verify accurate performance in qRT-PCR and improve your results!
Neil leblanc f7 rapid methods call amsterdam may 2010sva-slu_oie-cc
The document summarizes a strategic meeting discussing the development and use of multiplex assays using Luminex technology. Methods allow the detection of up to 100 unique assays in a single sample, and have been used to detect and subtype pestiviruses, avian influenza viruses, African swine fever virus, and cytokines. The technology provides a sensitive and specific platform for pathogen identification and characterization applicable to diagnostic laboratories.
RT-PCR is a sensitive technique for detecting and quantifying mRNA. It uses reverse transcriptase to synthesize cDNA from an RNA template, which is then used as a template for PCR amplification using DNA polymerase. RT-PCR can be performed as a one-step or two-step process and is commonly used in clinical microbiology labs to detect RNA viruses from specimens. The COVID-19 RT-PCR test analyzes respiratory specimens for SARS-CoV-2 RNA by amplifying small amounts into DNA to accurately diagnose infections.
This document provides information on several kits from Norgen Biotek Corp. for detecting HIV and related pathogens. It summarizes 3 kits that detect HIV: 1) an HIV quantitative RT-PCR kit that detects HIV RNA and has a reported linear range of 102-106 copies/μL, 2) an HIV proviral DNA PCR kit that detects HIV DNA with a linear range of 102-106 copies/μL, and 3) a plasma/serum HIV RT-PCR kit that isolates RNA from plasma/serum and detects HIV with a linear range of 10-8x106 VP/μL. It also summarizes kits for detecting Cryptococcus neoformans DNA and Pneumocystis jirove
Dr. Jeff Baxter - Lab Testing StandardizationJohn Blue
This document summarizes a study on standardizing Trichomonas foetus DNA testing across multiple laboratories. The study aimed to minimize variables that could influence sample or testing quality. It evaluated pooling of positive samples, different sample preparation methods, and real-time PCR protocols across five feeder labs compared to a central study lab. The study found 95.6% agreement between labs and confirmed 175 of 176 positive samples as T. foetus by DNA sequencing. Pooling was found to potentially miss some positives, with 1:5 pooling missing 4% and 1:3 pooling missing 3.5% of positives. The study supports standardizing sample collection, handling, preparation and PCR analysis to increase testing accuracy and consistency.
Quality Control of RNA Samples - For Gene-Expression Results you Can Rely onQIAGEN
By their very nature RNA molecules, especially mRNA and regulator RNA, are labile and can be highly unstable and sensitive to heat, UV and RNase contamination. The quality, relevance and scientific impact of gene expression results directly depends on the ability to extract RNA without losing any fraction of interest, while preserving the integrity of the biological information it carries. RNA quality control is thus critical to ensure high-quality results and for turning these results into actionable insights with confidence.
In this webinar, we will introduce you to the main parameters influencing RNA-based assays and their respective impact on downstream applications, discuss how to monitor them and cover the advantages of automation for quality control along complex workflows.
Quality Control of RNA Samples - For Gene-Expression Results you Can Rely on
Similar to A field-deployable automatic nucleic acid extraction and insulated isothermal RT-PCR system for sensitive on-site detection of avian influenza A virus
A panel was developed using the OpenArray platform to profile common respiratory tract pathogens via PCR. Assays were designed to target viral and bacterial sequences with high specificity and strain coverage. The panel demonstrated high specificity when tested against genomic standards. Pre-amplification improved sensitivity by enhancing detection of low copy targets. The panel provides a customizable and high-throughput tool for respiratory infection research.
Participants of the workshop learn the necessary background information and techniques to diagnose Sars-CoV-2 using the mobile diagnostic laboratory. The laboratory is shipped ready to use with all devices, reagents, certificates, and protocols. After one day of preparation together with a local assistant, a five-day course is given where every step is carried out by each participant. Experts accompany the learning process with written teaching materials, video training, virtual live coaching, and short exams to verify the learned content.
1) The document provides guidance on interpreting results from real-time reverse transcription polymerase chain reaction (rRT-PCR) diagnostic tests for avian influenza virus (AIV) and Newcastle disease virus (NDV).
2) Key steps in results interpretation include checking controls, evaluating growth curves, and recording cycle threshold (Ct) values. Suspect samples may require additional testing for confirmation.
3) Equivalency testing was performed between several real-time PCR instruments and chemistries. While results can vary, the Cepheid SmartCycler, ABI 7500, and Stratagene MX3005P were generally equivalent for AIV and NDV detection when using the same chemistry.
This document discusses avian influenza (H5N1) detection and analysis using real-time PCR. It describes the influenza virus, including its structure and types. Avian influenza is highly contagious in birds and can be fatal. Diagnosis involves virus isolation, serology tests, and molecular tests like RT-PCR and real-time RT-PCR. Real-time PCR allows for amplification and detection of targeted DNA sequences in one step and provides amplification curves and results in about 3 hours. The workflow described screens samples for influenza A using a matrix PCR, then checks positive samples for H5 and N1 subtypes using multiplex real-time PCR on the LightCycler system.
This document discusses practical approaches for diagnosing viral diseases in poultry, including clinical diagnosis, rapid field diagnostic tests, serological diagnosis, molecular diagnosis, and isolation/characterization. Clinical diagnosis is based on case history, clinical signs, examination of live/dead birds, and gross lesions. Rapid field tests can detect viruses but require high viral titers. Serological tests detect antibodies but have delays. Molecular diagnosis using PCR technologies can sensitively and specifically detect pathogens. The document emphasizes that clinical signs alone are not confirmatory and that multiple diagnostic approaches should be used to accurately diagnose poultry viral diseases.
The document describes the EpiTect ChIP qPCR System, a complete solution for chromatin immunoprecipitation (ChIP) followed by quantitative PCR (qPCR) analysis. The system includes optimized kits and reagents to simplify the multi-step ChIP workflow from chromatin extraction and immunoprecipitation to qPCR detection and data analysis. Key components are the EpiTect One-Day ChIP kit for streamlining the ChIP protocol, validated ChIP-grade antibody kits, EpiTect qPCR arrays containing pre-designed primers targeting promoter regions, and software for analyzing qPCR data. The system aims to remove technical challenges and allow researchers to focus on biological questions regarding protein-DNA interactions and epigenetic gene regulation
This document discusses transfusion-transmitted infections (TTIs) and methods for screening donated blood. It notes that TTIs include viruses like HIV, HBV, HCV that can remain undetected in the blood donor but be transmissible. Screening methods include serological tests like ELISA, CLIA, rapid tests, as well as nucleic acid amplification tests (NAATs) that can detect infections earlier. Implementing individual donor NAT in addition to serological screening provides an additional safety layer and reduces the risk window period for TTIs in blood donations.
S. Delannoy - IDENTYPATH: The genomic platform of ANSES for molecular detecti...EuFMD
This document discusses using microfluidic chips and high-throughput quantitative PCR (qPCR) to detect and type pathogens. It describes how the Fluidigm Biomark system allows processing thousands of qPCR reactions using nanoliter volumes across multiple samples and assays. Pre-amplification is also discussed as a way to increase sensitivity by amplifying targets before qPCR. The application of this approach for screening food, environmental, clinical, and bio-threat samples for viruses and bacteria is highlighted. In conclusion, it is suggested that high-throughput qPCR provides a valuable and flexible approach for large-scale pathogen screening.
The ChIP-qPCR assays provide pre-designed and validated real-time PCR primer assays that measure genomic DNA enrichment within chromatin immunoprecipitation samples. They save researchers time and money by eliminating the need to design their own assays. The assays provide quick, easy, and quantitative analysis of multiple promoter regions from a single ChIP sample using real-time PCR, addressing challenges of traditional ChIP workflows. The assays offer comprehensive coverage of human, mouse, and rat genomes and can be customized, allowing researchers to expand their ChIP experiments.
There are multiple HIV tests that can be used in combination as testing strategies. Testing strategies should use combinations of highly sensitive screening tests followed by more specific supplemental tests to accurately diagnose HIV status. Maintaining consistent testing strategies is important to ensure reliable results and proper clinical management of patients.
Pcr technology and its importance in covid 19 pandemicAnupam Maity
Since the discovery of the PCR technology, its application in the various fields is increased gradually. Based on to this principle, many variations of the PCR have been established. Year by year, it is upgraded very much. It is established as a most common and accurate technique for the detection of the various diseases in the field of medicine. Now it is a ‘Gold standard’ for the detection of covid-19 also, which is much needed to contain the spread of the virus. Though various detection techniques are there for detection, but real time RT-PCR (variation of PCR) is most reliable. Viral detection is based on a simple principle of nucleic acid (viral) amplification. Various manufacturing companies are manufacturing the PCR instrument. Though the accuracy of the instruments are slightly differ to each other.
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This document presents a next-generation sequencing (NGS)-based method for diagnosing COVID-19 using a single-step RNA extraction. The method was tested on 336 clinical samples and showed high accuracy compared to RT-PCR, the gold standard method. Key advantages of the NGS approach include higher scalability allowing thousands of samples to be tested simultaneously, and higher sensitivity to detect SARS-CoV-2 RNA compared to RT-PCR. The method provides individual qualitative results for each patient sample along with viral load estimates based on sequencing coverage.
This document provides information about antibody validation reviews from St. John's Laboratory. It details their process for scientists to receive free samples of antibodies and submit independent reviews. Reviews provide experimental results for specific antibodies used for techniques like western blot, immunohistochemistry, and immunofluorescence on various cell lines and tissues. Overall, the reviews demonstrate that the antibodies generated the expected target bands or labeling across different validation experiments.
Foregene's detection solution to covid 19Maggie Ma
In response to the Covid-19, Foregene developes the RT-PCR kit within 3 days.Based on Direct PCR tech, test centers needn't buy extra nucleic acid extraction kit and machine, just do PCR directly.It's so economical way. This kit is CE certificated, and welcomed by 10+ countries with stable quality and competitive prices.
The variant nucleic acid detection kit for Brazil, UK,India,and South Africa is also available with high specificity and sensitivity.
Are you a medical device importer?
Welcome your enquiry. E-mail:maggie@foregene.com
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3) Antibody tests detect antibodies produced after infection but cannot be used for diagnosis as they become positive later.
4) Newer modalities under research include multiplex assays, LAMP, CRISPR and other techniques for faster, portable, and higher-throughput Covid-19 testing.
This document describes pathway-powered PCR arrays for gene expression analysis. It discusses:
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Similar to A field-deployable automatic nucleic acid extraction and insulated isothermal RT-PCR system for sensitive on-site detection of avian influenza A virus (20)
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Here are some key objectives of communication with children:
Build Trust and Security:
Establish a safe and supportive environment where children feel comfortable expressing themselves.
Encourage Expression:
Enable children to articulate their thoughts, feelings, and experiences.
Promote Emotional Understanding:
Help children identify and understand their own emotions and the emotions of others.
Enhance Listening Skills:
Develop children’s ability to listen attentively and respond appropriately.
Foster Positive Relationships:
Strengthen the bond between children and caregivers, peers, and other adults.
Support Learning and Development:
Aid cognitive and language development through engaging and meaningful conversations.
Teach Social Skills:
Encourage polite, respectful, and empathetic interactions with others.
Resolve Conflicts:
Provide tools and guidance for children to handle disagreements constructively.
Encourage Independence:
Support children in making decisions and solving problems on their own.
Provide Reassurance and Comfort:
Offer comfort and understanding during times of distress or uncertainty.
Reinforce Positive Behavior:
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This particular slides consist of- what is hypotension,what are it's causes and it's effect on body, risk factors, symptoms,complications, diagnosis and role of physiotherapy in it.
This slide is very helpful for physiotherapy students and also for other medical and healthcare students.
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Hypotension, or low blood pressure, is when the pressure of blood circulating in the body is lower than normal or expected. It's only a problem if it negatively impacts the body and causes symptoms. Normal blood pressure is usually between 90/60 mmHg and 120/80 mmHg, but pressures below 90/60 are generally considered hypotensive.
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Here is a summary of Pneumothorax:
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VEDANTA AIR AMBULANCE SERVICES IN REWA AT A COST-EFFECTIVE PRICE.pdfVedanta A
Air Ambulance Services In Rewa works in close coordination with ground-based emergency services, including local Emergency Medical Services, fire departments, and law enforcement agencies.
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Emotional and Behavioural Problems in Children - Counselling and Family Thera...PsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
Hypertension and it's role of physiotherapy in it.Vishal kr Thakur
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NURSING MANAGEMENT OF PATIENT WITH EMPHYSEMA .PPTblessyjannu21
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Digital India will need a big trained army of Health Informatics educated & trained manpower in India.
Presently, generalist IT manpower does most of the work in the healthcare industry in India. Academic Health Informatics education is not readily available at school & health university level or IT education institutions in India.
We look into the evolution of health informatics and its applications in the healthcare industry.
HIMMS TIGER resources are available to assist Health Informatics education.
Indian Health universities, IT Education institutions, and the healthcare industry must proactively collaborate to start health informatics courses on a big scale. An advocacy push from various stakeholders is also needed for this goal.
Health informatics has huge employment potential and provides a big business opportunity for the healthcare industry. A big pool of trained health informatics manpower can lead to product & service innovations on a global scale in India.
Digital Health in India_Health Informatics Trained Manpower _DrDevTaneja_15.0...
A field-deployable automatic nucleic acid extraction and insulated isothermal RT-PCR system for sensitive on-site detection of avian influenza A virus
1. GeneReach Biotechnology Corporation
A field-deployable automatic nucleic acid
extraction and insulated isothermal RT-PCR
system for sensitive on-site detection of
avian influenza A virus
Simon Chung
GeneReach Biotechnology Corp.
Taiwan
4. GeneReach Biotechnology Corporation
Pen-side / Point of care PCR
• Needs
– Quick results for quick response / control measures / treatment
– Screening test in the field
• Challenge of the current system
– Takes time for transportation of samples from field to lab
• Requirements
– Easy to use: does not require high skill to operate
– Portable equipment (battery operated)
– Easy reagent storage (does not require freezers)
– Cost: inexpensive
– Equivalent level of sensitivity with qPCR in the lab = no compromise
5. GeneReach Biotechnology Corporation
Testing procedure of Pen-side PCR (tacomini and Pockit)
RNA extraction by Taco Mini PCR by Pockit
1. Open cover of pre-loaded plate
2. Add 200ul of sample to the first row
3. Set plate to tacomini and run
4. Wait for 25 minutes
5. Collect 150ul of RNA in elution buffer from the
last row to 1.5 ml tube
1. Take out pre-aliquoted reagent tubes
2. Add 50ul of reaction buffer to each tube
3. Add 5ul of RNA solution
4. Transfer 55ul from reagent tube to reaction tube
5. Set reaction tubes to Pockit and run
6. Wait for 40 minutes
7. Read results (positive / negative)
6. GeneReach Biotechnology Corporation
Materials and methods
• Avian influenza A virus (AIAV)-positive oropharyngeal swab (n =
26) and AIAV-spiked tissue (n = 8; brain, lung and spleen) were
subjected to extraction by tacoTM (tacoTM DNA/RNA Extraction
Kit) and RNeasy Mini Kit (Qiagen) simultaneously. Supernatant
alliquots of tissue homogenate were spiked with serial
dilutions of AIAV H5N1 strain (16A59). Reproducibility was
evaluated by triplicate extractions with 6 tissues and 6 swabs
of various AIAV titers. AIAV RNA from the samples were
quantified by a published qRT-PCR.
7. GeneReach Biotechnology Corporation
Material and methods (cont.)
• Analytical sensitivity of the RT-iiPCR was compared with a qRT-
PCR using an H5N1 isolate (16A59) or H7N9 isolate. The
inclusivity panel included H3, H4, H5, H6, H9 and H10 IAV
subtypes. NA from oropharyngeal swabs and tissue
homogenates (brain, lung and spleen) extracted by
tacoTM were tested to evaluate the RT-iiPCR by comparison
with the qRT-PCR. Interrater agreement was calculated by the
kappa test.
9. GeneReach Biotechnology Corporation
Test
Test 1: Test results with H7N9 virus diluted by 10-fold
Virus
dilution
-3 + + + 25.4 25.3 25.4
-4 + + + 28.7 28.6 28.5
-5 + + + 31.8 32.1 32.2
-6 + + + 35.3 35.3 36.2
-7 + + + 40.0 37.0 39.9
-8 + - - neg neg neg
Pockit M qPCR at Lab (Ct value)
Sensitivity and specificity of POCKIT Influenza A reagent for
Influenza A detection
Positive Negative
Pos 23 0 23
Neg 1 18 19
24 18 42
Specificity (%)=
Sensitivity & specificity of Pockit M compared with qPCR M
100.0
94.7
Pockit M
qPCR M*
Total
Total
Sensitivity (%) =
10. GeneReach Biotechnology Corporation
Sensitivity and specificity of POCKIT H7 for H7N9 detection
Test 2: Results of Pockit H7 with samples of Ct value between 27-37
qPCR*
Pos Neg Tested
27 1 0 1 100
28 2 0 2 100
29 2 0 2 100
30 1 0 1 100
31 3 0 3 100
32 2 0 2 100
33 2 0 2 100
34 3 0 3 100
35 3 1 4 75
36 1 0 1 100
37 0 1 1 0
Negative 0 8 8 100
Total 20 10 30
Pockit H7
% Agreement
Number of samples
Ct value
Test
Test 1: Test results with H7N9 virus diluted by 10-fold
Virus
dilution
-3 + + + 25.0 24.8 24.8
-4 + + + 28.2 28.1 28.1
-5 + + + 31.5 32.1 31.7
-6 + + + 35.3 34.8 36.1
-7 + - - 38.6 38.8 37.4
-8 - - - neg neg neg
Pockit H7 qPCR at Lab (Ct value)
Positive Negative
Pos 20 1 21
Neg 0 9 9
20 10 30
Total
Sensitivity (%) = 95.2
Specificity (%)= 100.0
Sensitivity & specificity of Pockit H7 compared with qPCR M
Pockit M
Total
qPCR M*
11. GeneReach Biotechnology Corporation
Pen-side PCR vs qPCR at Laboratory
Test Live bird market / Vet station Laboratory
Step 1:
RNA
extraction
Step 2:
Real-time
PCR
Portable equipment
Easy reagent storage (RT to 4C)
Simple test procedure
Simple result interpretation
Heavy equipment
Reagent stored in freezer
Requires skilled lab operator in
testing and interpretation
12. GeneReach Biotechnology Corporation
Advantages/limitations of tacomini/POCKIT
• Advantages
– Can detect influenza A virus at high sensitivity (100%) and specificity (95%)
– Can detect subtype H7 virus at high sensitivity (95%) and specificity (100%)
– Easy to operate: does not require high skill
– Kit is provided as ready-to-use: less chance of mistake
– Easy to carry: portable equipment, battery operated
– Low cost: POCKIT Micro Plus, tacomini
– Does not require high biosafety when samples are collected to Sample
storage buffer
• Limitations
– POCKIT H7 detect most H7 virus of Eurasian lineage, and not specific to HPAI
H7N9 (AIV HPAI H7 detection reagent has been validated in November)
– It is qualitative (Pos/Neg), not quantitative