This document provides information on techniques and procedures for serological tests. It discusses the necessary materials, including glassware, incubators, and rotating machines. It describes how to collect, prepare, and preserve serum, plasma, and cerebrospinal fluid specimens. Proper collection involves centrifugation to separate serum or plasma from whole blood. Specimens should be shipped promptly if not tested immediately. The document also covers complement inactivation through heating serum to prevent interference, and dilution techniques including serial dilution used to determine antibody titers.
This document provides information on techniques for serological tests. It discusses the materials needed, including glassware, incubators, and rotating machines. It describes how to collect, prepare, and preserve serological specimens like serum, plasma and cerebrospinal fluid. Proper shipment of specimens is also covered. The document explains complement inactivation through heating serum to prevent interference. Dilution techniques like serial dilution are outlined, along with calculating titers from dilution endpoints.
This document provides an overview of common biochemical tests used to differentiate bacterial species, including the catalase test, coagulase test, DNase test, bile solubility test, and litmus milk decolorization test. For each test, the document describes the principle, required materials, procedure, expected results, and positive and negative control organisms. The overall purpose is to teach medical laboratory science students how to perform and interpret the results of various biochemical tests to identify unknown bacterial isolates.
This document provides information on a human orosomucoid 2 (ORM2) ELISA kit that allows for the quantitative determination of ORM2 concentrations in biological samples like serum, plasma, tissue homogenates, and cell culture supernatants. It describes the intended use, test principle, materials included in the kit, sample collection and storage recommendations, limitations of the procedure, reagent preparation instructions, and the assay procedure.
This document provides guidelines for proper blood collection and handling of clinical specimens. It describes the steps for venipuncture including identifying the patient, selecting a vein, and collecting samples in various tubes according to order of draw. It also discusses collection and preservation of other specimens like sputum, urine, stool and CSF. The aim is to ensure samples are collected, transported and processed correctly for accurate diagnostic testing.
This document describes the microbiological assay method for quantifying antibiotics. It involves preparing standard solutions of known antibiotic concentrations and measuring their zones of inhibition against a test microbe. An unknown sample is also tested and its zone of inhibition is compared to the standard curve to determine its concentration. The key steps are:
1) Preparing standard solutions in increasing concentrations and an unknown at a median level.
2) Inoculating agar plates with a test microbe and creating wells for the solutions.
3) Measuring the zones of inhibition and calculating averages for the standards.
4) Comparing the unknown's zone of inhibition to the standard curve to determine its concentration.
This document discusses cytology techniques used to examine cells from body fluids and tissues. It describes how body fluids are collected and transported to the laboratory for processing. Common processing techniques include direct smears, centrifugation, cytocentrifugation, cell blocks and liquid-based preparations. Cell blocks allow cell pellets to be embedded in paraffin for sectioning, staining, and diagnostic evaluation like histology samples. This increases the sensitivity and specificity of cytology exams by enabling additional ancillary testing. The document provides details on various cell block preparation methods and their advantages.
This document discusses cytopreparatory techniques used in cytopathology. It covers different types of cytology samples like exfoliative cytology obtained from washing, smearing or brushing epithelial surfaces, as well as aspiration cytology using fine needle aspiration. The advantages of cytopathology are described as being non-invasive, allowing for faster reporting to guide clinicians, and being relatively inexpensive. The key steps in cytopreparatory techniques are outlined as specimen evaluation, smear preparation, fixation, and staining. Different fixation methods are also summarized.
This document discusses cytopreparatory techniques for cytology samples. It describes the different types of cytology samples like exfoliative cytology, aspiration cytology, and body fluids. The key steps in cytopreparatory techniques are outlined as evaluation of specimens, preparation of smears, fixation, and staining. Factors that can affect optimal cytological preparation like quality of specimen, fixative used, and stains are also summarized. Different fixation techniques including dry, wet, liquid-based, and lysing fixation for bloody samples are explained.
This document provides information on techniques for serological tests. It discusses the materials needed, including glassware, incubators, and rotating machines. It describes how to collect, prepare, and preserve serological specimens like serum, plasma and cerebrospinal fluid. Proper shipment of specimens is also covered. The document explains complement inactivation through heating serum to prevent interference. Dilution techniques like serial dilution are outlined, along with calculating titers from dilution endpoints.
This document provides an overview of common biochemical tests used to differentiate bacterial species, including the catalase test, coagulase test, DNase test, bile solubility test, and litmus milk decolorization test. For each test, the document describes the principle, required materials, procedure, expected results, and positive and negative control organisms. The overall purpose is to teach medical laboratory science students how to perform and interpret the results of various biochemical tests to identify unknown bacterial isolates.
This document provides information on a human orosomucoid 2 (ORM2) ELISA kit that allows for the quantitative determination of ORM2 concentrations in biological samples like serum, plasma, tissue homogenates, and cell culture supernatants. It describes the intended use, test principle, materials included in the kit, sample collection and storage recommendations, limitations of the procedure, reagent preparation instructions, and the assay procedure.
This document provides guidelines for proper blood collection and handling of clinical specimens. It describes the steps for venipuncture including identifying the patient, selecting a vein, and collecting samples in various tubes according to order of draw. It also discusses collection and preservation of other specimens like sputum, urine, stool and CSF. The aim is to ensure samples are collected, transported and processed correctly for accurate diagnostic testing.
This document describes the microbiological assay method for quantifying antibiotics. It involves preparing standard solutions of known antibiotic concentrations and measuring their zones of inhibition against a test microbe. An unknown sample is also tested and its zone of inhibition is compared to the standard curve to determine its concentration. The key steps are:
1) Preparing standard solutions in increasing concentrations and an unknown at a median level.
2) Inoculating agar plates with a test microbe and creating wells for the solutions.
3) Measuring the zones of inhibition and calculating averages for the standards.
4) Comparing the unknown's zone of inhibition to the standard curve to determine its concentration.
This document discusses cytology techniques used to examine cells from body fluids and tissues. It describes how body fluids are collected and transported to the laboratory for processing. Common processing techniques include direct smears, centrifugation, cytocentrifugation, cell blocks and liquid-based preparations. Cell blocks allow cell pellets to be embedded in paraffin for sectioning, staining, and diagnostic evaluation like histology samples. This increases the sensitivity and specificity of cytology exams by enabling additional ancillary testing. The document provides details on various cell block preparation methods and their advantages.
This document discusses cytopreparatory techniques used in cytopathology. It covers different types of cytology samples like exfoliative cytology obtained from washing, smearing or brushing epithelial surfaces, as well as aspiration cytology using fine needle aspiration. The advantages of cytopathology are described as being non-invasive, allowing for faster reporting to guide clinicians, and being relatively inexpensive. The key steps in cytopreparatory techniques are outlined as specimen evaluation, smear preparation, fixation, and staining. Different fixation methods are also summarized.
This document discusses cytopreparatory techniques for cytology samples. It describes the different types of cytology samples like exfoliative cytology, aspiration cytology, and body fluids. The key steps in cytopreparatory techniques are outlined as evaluation of specimens, preparation of smears, fixation, and staining. Factors that can affect optimal cytological preparation like quality of specimen, fixative used, and stains are also summarized. Different fixation techniques including dry, wet, liquid-based, and lysing fixation for bloody samples are explained.
The document discusses ABO blood grouping methods and procedures. The two main methods are the slide method and spin tube method. The slide method uses glass slides while the spin tube method uses test tubes. Procedures include preparing red blood cell suspensions, adding blood and antisera to slides or tubes, incubating, and observing for agglutination. Quality control and potential sources of error are also outlined.
Sample preparation involves collecting and processing samples to isolate proteins, DNA, or other molecules of interest. For protein samples, cells or tissues are lysed to release proteins, which are then separated from other components through techniques like centrifugation or filtration. Purification methods separate protein mixtures based on properties like size, charge, or solubility using chromatography, electrophoresis, or precipitation. DNA extraction from blood involves lysing cells, precipitating DNA, and purifying it from other cell components. Proper sample preparation is essential for downstream diagnostic and analytical tests.
This document provides information on various blood banking and laboratory procedures. It discusses professional, replacement, and voluntary blood donors. It also describes agglutination reactions, blood grouping procedures using slide agglutination, blood component preparation including red blood cells, platelets, and plasma, cross matching methods, hemoglobin estimation techniques, malaria parasite testing, and pH meter principles and use.
Washed red blood cell suspensions are prepared to remove plasma proteins that could interfere with antigen-antibody reactions during blood typing tests. The red blood cells are separated from whole blood via centrifugation and washed with saline to remove plasma. This helps remove soluble antigens, interfering proteins, and substances that could cause false positive reactions. The washed red blood cells are then suspended in saline at a 3-5% concentration for use in blood typing tests.
1. The document describes procedures for titrating anti-D antibodies, performing Du testing, cross matching donor and patient blood, and direct and indirect Coombs testing.
2. Key steps for titration include serially diluting a test serum, adding diluted serum to test tubes containing red blood cells, and determining the titer by the highest dilution showing agglutination.
3. Du testing, cross matching, and Coombs testing involve incubating patient and donor blood components together, then checking for agglutination with or without the addition of anti-globulin reagent to indicate blood compatibility.
1) A blood bank stores and tests donated blood for use in transfusions. It classifies blood into four main groups (A, B, AB, O) based on the presence of antigens on red blood cells.
2) Testing done in blood banks includes preparing red blood cell suspensions, performing blood grouping using slide and tube methods, reverse grouping, weak D testing to identify weakly positive Rh types, and cross matching donor blood with recipient serum to check for compatibility.
3) Donor criteria include being age 18-60, weighing over 55kg, and being free of transfusion-transmissible infections and diseases. Certain tests and standards are followed to ensure safe collection and storage of blood for transfusions.
The document discusses quality assurance in haemostasis laboratory testing. It outlines the importance of accuracy and precision in test results. There are three main types of quality assurance: internal quality control, external quality control, and participation in proficiency testing programs. The document also describes pre-analytic, analytic, and post-analytic factors that can affect test results and outlines standard operating procedures to ensure quality at each stage of testing. Maintaining reliable test results requires strict control of all variables from specimen collection through analysis and reporting.
Adaptive immunity can be acquired naturally through infection or artificially through immunoglobulin injection or vaccination. It involves two main mechanisms: humoral immunity mediated by antibodies produced by B lymphocytes, and cell-mediated immunity mediated by T lymphocytes. Adaptive immunity is characterized by antigen specificity, diversity, immunological memory, self/non-self discrimination, and anamnestic responses. B and T lymphocytes develop and mature in the bone marrow and thymus, respectively, undergoing selection processes to respond appropriately to pathogens.
Antigens are molecules that can trigger an immune response. The immunogenicity or ability of an antigen to provoke an immune response is influenced by several factors including the antigen's size, dose, route of administration, purity, and whether the antigen is attached to a carrier protein. The immune system has developed mechanisms to discriminate between self and foreign antigens to avoid autoimmune responses.
The document discusses adaptive immunity and the immune response. It describes the four phases of the adaptive immune response as encounter, activation, attack, and memory. Acquired immunity develops over one's lifetime from vaccines, infections, or antibodies from others. The adaptive immune response involves B cells, helper T cells and cytotoxic T cells. It also discusses the specificity, adaptiveness, discrimination between self and non-self, and memory properties of the adaptive immune response. The major cell types involved are T helper cells, cytotoxic T cells, T memory cells, and regulatory T cells. The roles and mechanisms of these cell types are described in detail.
Adaptive immunity can be acquired naturally through infection or artificially through immunoglobulin injection or vaccination. It involves two main mechanisms: humoral immunity where B cells produce antibodies, and cell-mediated immunity where T cells attack foreign material directly or through cytokine release. Adaptive immunity is characterized by antigen specificity, diversity, immunological memory, self/non-self recognition, and anamnestic responses. B and T cells develop through selection processes in the bone marrow and thymus respectively to respond appropriately to pathogens.
Professional antigen presenting cells such as macrophages, dendritic cells, and B cells express MHC class II molecules. These cells uptake antigens through phagocytosis, pinocytosis, or receptor-mediated endocytosis. The antigens are then processed through two pathways: the MHC class I pathway processes endogenous antigens in the cytosol using the proteasome, while the MHC class II pathway processes exogenous antigens in lysosomes. The processed antigen peptides bind to MHC molecules, which are then presented on the cell surface to activate CD4+ or CD8+ T cells depending on the MHC molecule.
The document discusses the components of the innate immune system. It describes the physical, chemical, biological, and physiological barriers that block pathogens from entering the body. It also explains the cellular responses involving phagocytes that destroy pathogens through phagocytosis. The complement system and inflammatory response are blood proteins and processes that help eliminate pathogens. Key cells of innate immunity discussed are neutrophils, macrophages, dendritic cells, and natural killer cells.
Cytokines are low molecular weight proteins that are important mediators of the immune system. They can be classified into interleukins, interferons, tumor necrosis factors, colony stimulating factors, chemokines, and growth factors. Cytokines act through specific cell surface receptors and have pleiotropic, redundant, synergistic and antagonistic effects. They are involved in innate immunity, adaptive immunity, inflammation, and hematopoiesis. Therapeutic uses of cytokines include treatment of viral infections, cancer, immunodeficiencies, and autoimmune diseases through administration of cytokines or anti-cytokine antibodies.
Immunoassays are biochemical methods that use the specificity of antigen-antibody reactions to detect and quantify target molecules in biological samples. There are several types of immunoassays including enzyme immunoassays (such as ELISA), radioimmunoassays, counting immunoassays, fluoroimmunoassays, and chemiluminescence immunoassays. ELISA is a common immunoassay that uses an enzyme-linked antibody to detect an antigen and produces a color change to quantify the target molecule.
Major Histocompatibility Complex & transplantation 3rd.pptxSherzadMajeed1
The major histocompatibility complex (MHC) plays a key role in the immune system's response to transplants and infections. The MHC is a set of genes that encode MHC molecules which are expressed on nearly all cells and present antigen peptides. There are three main classes of MHC molecules - Class I presents antigens to CD8+ T cells, Class II presents antigens to CD4+ T cells, and Class III encodes proteins involved in immune processes. MHC molecules are important in transplant rejection, as mismatches can trigger an immune response against the donor tissue. Careful HLA typing and matching is done to minimize rejection.
Molecular Immunopatological Study of ACE2 Expression and Polymorphism.pptxSherzadMajeed1
This document summarizes the progress of the author's Ph.D. research from June 2021 to January 2023. The goal of the research is to examine the impacts of Covid-19 on hospitalized patients in Duhok province, Kurdistan region, by analyzing how SARS-CoV-2 induces cytokine storms and oxidative stress that increase fatality rates. It also evaluates the effects of multiple drug therapies on hospitalized patients. The study will analyze whole genome sequencing and phylogeny of SARS-CoV-2 cases, examine ACE2 expression rates in different age groups and genders and their link to infection severity, and investigate the interaction between the SARS-CoV-2 spike protein and ACE2-expressing cells
Class I and class II MHC molecules bind antigenic peptides derived from degraded antigens. MHC molecules present antigen peptides to T cells but do not have the fine specificity of antibodies or T cell receptors. The distal regions of MHC molecules display allelic variation that results in different antigen-binding clefts with varying specificities. For an antigen to be recognized by T cells, it must be degraded into peptides that form complexes with class I or class II MHC molecules on the cell surface in a process called antigen processing and presentation.
The innate immune system provides the first line of defense against pathogens through physical, chemical, and cellular barriers. Physical barriers include the skin and mucous membranes. Chemical barriers secrete antimicrobial peptides. Cellular responses involve phagocytes such as macrophages and neutrophils that engulf and destroy pathogens. Natural killer cells attack virus-infected cells. Mast cells and dendritic cells help initiate inflammation and adaptive immune responses. The innate system provides broad but non-specific protection against microbes.
Whole genome sequencing was performed on SARS-CoV-2 samples isolated from COVID-19 patients in Duhok, Iraq. 36 samples were sequenced and their genomes were analyzed and compared to reference genomes. A total of 39 mutations were identified within the genomes, with 27 being non-synonymous single nucleotide variants that resulted in amino acid substitutions in the encoded proteins. The mutations were submitted to public databases to help track viral evolution.
The Pan India 1000 SARS-CoV-2 Genome Sequencing Consortium sequenced 1000 viral genomes from COVID-19 patient samples collected across India. Initial results found multiple SARS-CoV-2 lineages circulating, including a predominance of the A2a haplotype containing the D614G mutation. Mutations were observed geographically clustered and in important viral genome regions. The sequencing data will help understand virus transmission and evolution in India to improve response efforts. The consortium was coordinated by the National Institute of Biomedical Genomics and involved teams from various institutes performing sequencing, analysis, and collaborating with clinical organizations.
The document discusses ABO blood grouping methods and procedures. The two main methods are the slide method and spin tube method. The slide method uses glass slides while the spin tube method uses test tubes. Procedures include preparing red blood cell suspensions, adding blood and antisera to slides or tubes, incubating, and observing for agglutination. Quality control and potential sources of error are also outlined.
Sample preparation involves collecting and processing samples to isolate proteins, DNA, or other molecules of interest. For protein samples, cells or tissues are lysed to release proteins, which are then separated from other components through techniques like centrifugation or filtration. Purification methods separate protein mixtures based on properties like size, charge, or solubility using chromatography, electrophoresis, or precipitation. DNA extraction from blood involves lysing cells, precipitating DNA, and purifying it from other cell components. Proper sample preparation is essential for downstream diagnostic and analytical tests.
This document provides information on various blood banking and laboratory procedures. It discusses professional, replacement, and voluntary blood donors. It also describes agglutination reactions, blood grouping procedures using slide agglutination, blood component preparation including red blood cells, platelets, and plasma, cross matching methods, hemoglobin estimation techniques, malaria parasite testing, and pH meter principles and use.
Washed red blood cell suspensions are prepared to remove plasma proteins that could interfere with antigen-antibody reactions during blood typing tests. The red blood cells are separated from whole blood via centrifugation and washed with saline to remove plasma. This helps remove soluble antigens, interfering proteins, and substances that could cause false positive reactions. The washed red blood cells are then suspended in saline at a 3-5% concentration for use in blood typing tests.
1. The document describes procedures for titrating anti-D antibodies, performing Du testing, cross matching donor and patient blood, and direct and indirect Coombs testing.
2. Key steps for titration include serially diluting a test serum, adding diluted serum to test tubes containing red blood cells, and determining the titer by the highest dilution showing agglutination.
3. Du testing, cross matching, and Coombs testing involve incubating patient and donor blood components together, then checking for agglutination with or without the addition of anti-globulin reagent to indicate blood compatibility.
1) A blood bank stores and tests donated blood for use in transfusions. It classifies blood into four main groups (A, B, AB, O) based on the presence of antigens on red blood cells.
2) Testing done in blood banks includes preparing red blood cell suspensions, performing blood grouping using slide and tube methods, reverse grouping, weak D testing to identify weakly positive Rh types, and cross matching donor blood with recipient serum to check for compatibility.
3) Donor criteria include being age 18-60, weighing over 55kg, and being free of transfusion-transmissible infections and diseases. Certain tests and standards are followed to ensure safe collection and storage of blood for transfusions.
The document discusses quality assurance in haemostasis laboratory testing. It outlines the importance of accuracy and precision in test results. There are three main types of quality assurance: internal quality control, external quality control, and participation in proficiency testing programs. The document also describes pre-analytic, analytic, and post-analytic factors that can affect test results and outlines standard operating procedures to ensure quality at each stage of testing. Maintaining reliable test results requires strict control of all variables from specimen collection through analysis and reporting.
Adaptive immunity can be acquired naturally through infection or artificially through immunoglobulin injection or vaccination. It involves two main mechanisms: humoral immunity mediated by antibodies produced by B lymphocytes, and cell-mediated immunity mediated by T lymphocytes. Adaptive immunity is characterized by antigen specificity, diversity, immunological memory, self/non-self discrimination, and anamnestic responses. B and T lymphocytes develop and mature in the bone marrow and thymus, respectively, undergoing selection processes to respond appropriately to pathogens.
Antigens are molecules that can trigger an immune response. The immunogenicity or ability of an antigen to provoke an immune response is influenced by several factors including the antigen's size, dose, route of administration, purity, and whether the antigen is attached to a carrier protein. The immune system has developed mechanisms to discriminate between self and foreign antigens to avoid autoimmune responses.
The document discusses adaptive immunity and the immune response. It describes the four phases of the adaptive immune response as encounter, activation, attack, and memory. Acquired immunity develops over one's lifetime from vaccines, infections, or antibodies from others. The adaptive immune response involves B cells, helper T cells and cytotoxic T cells. It also discusses the specificity, adaptiveness, discrimination between self and non-self, and memory properties of the adaptive immune response. The major cell types involved are T helper cells, cytotoxic T cells, T memory cells, and regulatory T cells. The roles and mechanisms of these cell types are described in detail.
Adaptive immunity can be acquired naturally through infection or artificially through immunoglobulin injection or vaccination. It involves two main mechanisms: humoral immunity where B cells produce antibodies, and cell-mediated immunity where T cells attack foreign material directly or through cytokine release. Adaptive immunity is characterized by antigen specificity, diversity, immunological memory, self/non-self recognition, and anamnestic responses. B and T cells develop through selection processes in the bone marrow and thymus respectively to respond appropriately to pathogens.
Professional antigen presenting cells such as macrophages, dendritic cells, and B cells express MHC class II molecules. These cells uptake antigens through phagocytosis, pinocytosis, or receptor-mediated endocytosis. The antigens are then processed through two pathways: the MHC class I pathway processes endogenous antigens in the cytosol using the proteasome, while the MHC class II pathway processes exogenous antigens in lysosomes. The processed antigen peptides bind to MHC molecules, which are then presented on the cell surface to activate CD4+ or CD8+ T cells depending on the MHC molecule.
The document discusses the components of the innate immune system. It describes the physical, chemical, biological, and physiological barriers that block pathogens from entering the body. It also explains the cellular responses involving phagocytes that destroy pathogens through phagocytosis. The complement system and inflammatory response are blood proteins and processes that help eliminate pathogens. Key cells of innate immunity discussed are neutrophils, macrophages, dendritic cells, and natural killer cells.
Cytokines are low molecular weight proteins that are important mediators of the immune system. They can be classified into interleukins, interferons, tumor necrosis factors, colony stimulating factors, chemokines, and growth factors. Cytokines act through specific cell surface receptors and have pleiotropic, redundant, synergistic and antagonistic effects. They are involved in innate immunity, adaptive immunity, inflammation, and hematopoiesis. Therapeutic uses of cytokines include treatment of viral infections, cancer, immunodeficiencies, and autoimmune diseases through administration of cytokines or anti-cytokine antibodies.
Immunoassays are biochemical methods that use the specificity of antigen-antibody reactions to detect and quantify target molecules in biological samples. There are several types of immunoassays including enzyme immunoassays (such as ELISA), radioimmunoassays, counting immunoassays, fluoroimmunoassays, and chemiluminescence immunoassays. ELISA is a common immunoassay that uses an enzyme-linked antibody to detect an antigen and produces a color change to quantify the target molecule.
Major Histocompatibility Complex & transplantation 3rd.pptxSherzadMajeed1
The major histocompatibility complex (MHC) plays a key role in the immune system's response to transplants and infections. The MHC is a set of genes that encode MHC molecules which are expressed on nearly all cells and present antigen peptides. There are three main classes of MHC molecules - Class I presents antigens to CD8+ T cells, Class II presents antigens to CD4+ T cells, and Class III encodes proteins involved in immune processes. MHC molecules are important in transplant rejection, as mismatches can trigger an immune response against the donor tissue. Careful HLA typing and matching is done to minimize rejection.
Molecular Immunopatological Study of ACE2 Expression and Polymorphism.pptxSherzadMajeed1
This document summarizes the progress of the author's Ph.D. research from June 2021 to January 2023. The goal of the research is to examine the impacts of Covid-19 on hospitalized patients in Duhok province, Kurdistan region, by analyzing how SARS-CoV-2 induces cytokine storms and oxidative stress that increase fatality rates. It also evaluates the effects of multiple drug therapies on hospitalized patients. The study will analyze whole genome sequencing and phylogeny of SARS-CoV-2 cases, examine ACE2 expression rates in different age groups and genders and their link to infection severity, and investigate the interaction between the SARS-CoV-2 spike protein and ACE2-expressing cells
Class I and class II MHC molecules bind antigenic peptides derived from degraded antigens. MHC molecules present antigen peptides to T cells but do not have the fine specificity of antibodies or T cell receptors. The distal regions of MHC molecules display allelic variation that results in different antigen-binding clefts with varying specificities. For an antigen to be recognized by T cells, it must be degraded into peptides that form complexes with class I or class II MHC molecules on the cell surface in a process called antigen processing and presentation.
The innate immune system provides the first line of defense against pathogens through physical, chemical, and cellular barriers. Physical barriers include the skin and mucous membranes. Chemical barriers secrete antimicrobial peptides. Cellular responses involve phagocytes such as macrophages and neutrophils that engulf and destroy pathogens. Natural killer cells attack virus-infected cells. Mast cells and dendritic cells help initiate inflammation and adaptive immune responses. The innate system provides broad but non-specific protection against microbes.
Whole genome sequencing was performed on SARS-CoV-2 samples isolated from COVID-19 patients in Duhok, Iraq. 36 samples were sequenced and their genomes were analyzed and compared to reference genomes. A total of 39 mutations were identified within the genomes, with 27 being non-synonymous single nucleotide variants that resulted in amino acid substitutions in the encoded proteins. The mutations were submitted to public databases to help track viral evolution.
The Pan India 1000 SARS-CoV-2 Genome Sequencing Consortium sequenced 1000 viral genomes from COVID-19 patient samples collected across India. Initial results found multiple SARS-CoV-2 lineages circulating, including a predominance of the A2a haplotype containing the D614G mutation. Mutations were observed geographically clustered and in important viral genome regions. The sequencing data will help understand virus transmission and evolution in India to improve response efforts. The consortium was coordinated by the National Institute of Biomedical Genomics and involved teams from various institutes performing sequencing, analysis, and collaborating with clinical organizations.
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share - Lions, tigers, AI and health misinformation, oh my!.pptxTina Purnat
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Does Over-Masturbation Contribute to Chronic Prostatitis.pptxwalterHu5
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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.
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2. Learning Objective
The students should be able to:
1. List material and equipment for serological tests
2. Collect, preserve and prepare serological specimens
3. Run complement inactivation procedure and state
its importance
4. Run serial dilution, determine end point and titer.
3. Outline
1. Introduction
2. Materials necessary for basic serologic tests
3. Collection, preparation and preservation of
serologic al tests
4. Shipment of serological specimens
5. Complement inactivation
6. Dilution and Serial dilution
4. 1. Introduction
Dilution is the act of making a weaker solution
from a strong solution.
Serial dilution The systematic re-dilution of a
fluid number of times is called a Serial dilution
Titer is the reciprocal of the highest dilution
showing a positive reaction
Complement is a group of non-immunoglobulin
plasma proteins that are sequentially activated by
Ag–Ab complexes
5. Types of glassware include:
Test tubes
Glass slides
Serological pipette with a size of 10ml, 5ml, 2ml
and 1ml.
2. Materials necessary for basic
serologic tests
6. 2. Materials necessary for basic
serologic tests
Glassware
Dirty glassware easily affects serological tests.
After using all the glassware (test tube, beaker,
pipette, etc) they should be soaked in detergent
for several hours and rinsed several times in tap
water.
Finally, allow drying by placing in a dry oven or
dust free place. Test tubes and pipettes should
not be scratched or broken, which will interfere
with the reading of a test.
8. Constant Temperature Device
Incubators and water baths are used in
serological tests. These materials are
electrically operated and have thermostat that
hold the temperature within the required
limits. These devices should be checked prior
to use by a thermometer.
2. Materials necessary for basic
serologic tests
9. Rotating Machine
Rotating machines are required to facilitate
antigen antibody reactions. Such machines have
a flat plate, which rotate at a prescribed rate of
speed. A knob located on the front of the
machine controls the number of revolutions per
minute.
2. Materials necessary for basic
serologic tests
10.
11. 3. Collection, Preparation And
Preservation Of Specimens
For Serologic tests
A.Types of Specimens
Specimens that are used for serologic test
include: serum, plasma and cerebrospinal
fluid.
Serum or plasma samples could be obtained from
venous blood, which can be collected by the
laboratory personnel.
CSF should be collected by a physician or
12. B. Serum or plasma sample collection
Collect 2-3ml of venous blood from a patient
using a sterile syringe and needle.
If serum is required, allow the whole blood to clot
at room temperature for at least one hour,
Centrifuge the clotted blood for 10 minutes at
2000 rpm.
3. Collection, Preparation And
Preservation Of Specimens
For Serologic tests
13. B. Serum or plasma sample collection
Transfer the serum to a labeled tube with a
paster pipette and rubber bulb.
Plasma samples are obtained by treating fresh
blood with anticoagulant,
Centrifuge and separate the supernatant.
3. Collection, Preparation And
Preservation Of Specimens
For Serologic tests
14. B. Serum or plasma sample collection
The specimen should be free from hemolyzed blood.
Finally, seal the specimen containing tube; the tube
should be labeled with full patient's identification
(age, sex, code number, etc).
The test should be performed within hours after
sample collection, if this could not be done preserve
it at -20oc.
2. Collection, Preparation And
Preservation Of Specimens
For Serologic tests
15. Most health center and clinic laboratories are
limited in the diagnostic procedures that can be
carried out and have to ship serologic specimens
to other laboratories.
Before shipment, the following things should be
considered.
Don't ship whole blood unless the tests to be
performed require whole blood.
Don't inactivate serum or plasma.
4. Shipment of serological
specimens
16. Serum, plasma, and CSF should be handled as
follows:
Collect and process specimens under sterile
conditions.
Ship specimens by the fastest route as soon after
collection as possible.
Don't ship whole blood unless the test to be performed
required whole blood.
Remove cells from plasma and clot from serum before
shipment.
4. Shipment of serological
specimens
17. Serum, plasma, and CSF should be handled as
follows:
Don't inactivate serum or plasma before mailing.
Keep the specimen and packing container in the
refrigerator until time of shipment.
Shipment is requires several days preserve by
refrigeration in transit. First, freeze the
specimen; then pack and ship in a well-insulated
container with dry ice.
4. Shipment of serological
specimens
18. Complement is a group of non-immunoglobulin
plasma proteins that are sequentially activated
by Ag–Ab complexes (or directly by microbial
constituents) and cause irreversible damage to
membrane of cellular target
5. Complement inactivation
19. Some tests need inactivated serum. Others do
not.
Inactivation may be important since complement
promotes lysis of erythrocytes and can contribute
to false test results in tests using RBCs.
Some complement components may also cause
false agglutination in some tests.
5. Complement inactivation
20. Complement components can be inactivated by
of three mechanism
Spontaneous decay
Enzymatic degradation of C4, C3 and C5
rapidly decay
Stoichiometric inhibition
5. Complement inactivation
21. The complement in serum must be inactivated
usually by stoichiometric inhibition for most
serological testing.
To inactivate complement, place tubes of serum
in hot water bath (56c) for 30min
If the protein complement is not inactivated it will
promote lysis of the red cells and other types of
cells and can therefore produce invalid results
5. Complement inactivation
22. Complement is also known to interfere with
certain tests for syphilis.
Serum samples to be tested more than 4 hours
after inactivation should be reheated at 560c for
10 minutes and allowed to cool to room
temperature
5. Complement inactivation
23. Dilution is the act of making a weaker solution
from a strong solution.
Adding a diluent such as water or saline,
which contains none of the material being
diluted, is used to do this.
6. Dilution
24. Dilution techniques
Dilutions can be used in the laboratory to change
the concentration of the body fluids, such as serum
so that it is consistent with the range of an assay.
Making dilutions can also be necessary to prepare
reagents and standards.
Dilution has two parts: diluents and solute.
6. Dilution
25. A dilution involves adding of a substance, the
diluent to other substances, the solute.
Dilutions show the relative amount of the solute
in the dilute solution.
It is an indicator of concentration, not volume.
6. Dilution
26. For a 1:10 dilution, the dilution factor is 10. For a :
b dilution the dilution factor is b.
6. Dilution
27. Technique
Two liquids of very different compositions (density,
or surface tension) is required
An exact volume of concentrated solute is added to
a calibrated flask or container, and then diluent is
added to the required volume.
Adequate mixing must take place to ensure
homogeneity
6. Dilution
28. E.g.,
if you want to prepare 1:10 dilution
Take 1 ml solute
Take 9 ml solvent
Then mix
1st
2nd
6. Dilution
29. When a solution is diluted with water, its
concentration is decreased and its volume is
increased. But the total amount of solute remains
constant.
Mathematical expressions of the dilutions are;
CiVi = CfVf Where, Ci is initial concentration
Vi is initial volume.
Cf final concentration
Vf is final volume.
6. Dilution
30. Serial dilutions
Serial dilutions are a unique type of dilution
techniques.
In serial dilution, all dilutions, except the 1st are
prepared from the previous dilution and all
dilutions made after the initial dilution are the
same.
6. Dilution
31. Serial dilutions are used to prepare sets of
standard solutions and are also used to
prepare patient's samples to analyze
components that can exist over a wide
concentration range, such as antibody titers.
6. Dilution
33. An example of the serial dilution is as follows: -
Into each of ten test tubes is measured 0.5 ml of
saline 1/2 ml of serum is placed in the 1st tube and
mixed.
Since there is 0.5 ml of serum in a total volume of
1.0 ml; a 0.5:1 or a 1:2 dilution exists in the first
tube.
6. Dilution
34. Now, 0.5 ml of this solution is removed and mixed
with the 0.5 ml of saline in the 2nd tube; this gives
another 1:2 dilution, but since the 0.5 ml of solution
put into the 2nd tube is already a 1:2 dilution of the
serum, the dilution of serum in the 2nd tube is one
half that of the 1st tube or 1/2 of ½ =1/4 or 1:4.
This and, by applying the above reasoning, the
dilutions of serum are found to be (1/2)10 = 1/1024
or 1: 1024 in the 10th tube.
6. Dilution
35. Try the following problems
For ASO titer, tube 1 contains 0.8ml 0f saline,
tubes 2 to 5 contain 0.5ml of saline; 0.2ml of
serum is added to tube 1, and serial dilutions
using 0.5ml are carried out in the remaining
tubes. What is the dilution in each tube?
Explain the shipment of specimen and
complement inactivation.
Review questions
36. Reference
1. Tizard. Immunology an introduction,4th edition
,Saunders publishing,1994
2. Naville J. Bryant Laboratory Immunology and
Serology 3rd edition. Serological services
Ltd.Toronto,Ontario,Canada,1992
3. Mary Louise .Immunology and Serology in
Laboratory medicine 3rd edition