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Indirect immunoperoxidase assay (IPA) is a laboratory technique used to detect and
Immunohistochemistry Protocols, Buffers and Troubleshooting
{{Immunologic techniques and tests, state expand Biochemistry methods Diagnostic virology Immunologic tests Laboratory techniques
Indirect immunoperoxidase assay (IPA) is a laboratory technique used to detect and titrate
Titration (also known as titrimetry and volumetric analysis) is a common laboratory method of quantitative chemical analysis to determine the concentration of an identified analyte (a substance to be analyzed). A reagent, termed the ''titrant'' ...
viruses that do not cause measurable cytopathic effect
Cytopathic effect or cytopathogenic effect (abbreviated CPE) refers to structural changes in host cells that are caused by viral invasion. The infecting virus causes lysis of the host cell or when the cell dies without lysis due to an inability to ...
s and cannot be measured by classical plaque assays. These viruses include human coronavirus 229E and OC43.
Methodology
Susceptible cells are inoculated with serial logarithmic dilutions of samples in a 96-well plate. After viral growth, viral detection by IPA yields the infectious virus titer, expressed as tissue culture infectious dose (TCID50). This represents the dilution of a virus-containing sample at which half of a series of laboratory wells contain replicating viruses. This technique is a reliable method for the titration of human coronaviruses (HCoV) in biological samples (cells, tissues, or fluids). It is also reliable in the detection of antibodies tohuman cytomegalovirus
''Human betaherpesvirus 5'', also called human cytomegalovirus (HCMV), is species of virus in the genus ''Cytomegalovirus'', which in turn is a member of the viral family known as ''Herpesviridae'' or herpesviruses. It is also commonly called ...
.G Gerna, C J McCloud and R W Chambers: Immunoperoxidase technique for detection of antibodies to Human cytomegalovirus. J. Clin. Microbiol. 1976, 3(3):364.
See also
*Coronavirus
Coronaviruses are a group of related RNA viruses that cause diseases in mammals and birds. In humans and birds, they cause respiratory tract infections that can range from mild to lethal. Mild illnesses in humans include some cases of the com ...
*Immunoassay
An immunoassay (IA) is a biochemical test that measures the presence or concentration of a macromolecule or a small molecule in a solution through the use of an antibody (usually) or an antigen (sometimes). The molecule detected by the immunoass ...
*Immunoperoxidase Immunoperoxidase is a type of immunostain used in molecular biology, medical research, and clinical diagnostics. In particular, immunoperoxidase reactions refer to a sub-class of immunohistochemical or immunocytochemical procedures in which the ant ...
*Virus quantification
Virus quantification involves counting the number of viruses in a specific volume to determine the virus concentration. It is used in both research and development (R&D) in commercial and academic laboratories as well as production situations whe ...
References
External links
*Immunohistochemistry Protocols, Buffers and Troubleshooting
{{Immunologic techniques and tests, state expand Biochemistry methods Diagnostic virology Immunologic tests Laboratory techniques