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A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Hepatitis C: an Overview of Various Laboratory Assays with their Mode of Diagnostic Cooperation
Insights
This review details hepatitis C virus (HCV) diagnostic assays, including antibody, core antigen, and RNA tests. Advanced enzyme-linked immunosorbent assays (ELISA) and RT-PCR have improved HCV diagnosis, reducing the need for older confirmatory methods.
Area of Science:
- Virology
- Immunology
- Diagnostic Medicine
Background:
- Hepatitis C virus (HCV) diagnosis has evolved significantly.
- Early diagnostic methods included antibody and core antigen assays.
- RNA detection assays provide crucial diagnostic information.
Purpose of the Study:
- To chronologically review diagnostic assays for HCV.
- To compare the utility of various HCV testing methods.
- To highlight advancements in HCV detection and diagnosis.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA) for antibodies and core antigen.
- Rapid diagnostic tests (RDTs) and microarray assays for HCV antibodies.
- Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) for HCV RNA detection.
Main Results:
- Third and fourth generation ELISA, alongside RT-PCR, offer sensitive HCV detection.
- Recombinant immunoblot assay (RIBA) is largely superseded by modern ELISA and RNA tests.
- ELISA and RIBA indicate past or present infection, while RT-PCR confirms active infection.
Conclusions:
- ELISA and RT-PCR are vital for accurate HCV diagnosis, differentiating acute, chronic, and false results.
- HCV genotyping and subtyping are essential for effective treatment strategies and epidemiological studies.
Background:
This review describes in chronological order the different assays for hepatitis C virus (HCV) antibodies, for the core antigen and for the HCV-RNA.
Methods:
By ascending chronological order, the enzyme-linked immunosorbent assay (ELISA), rapid diagnostic tests (RDTs), HCV-Ab IgG avidity index (HCV AI), and Cy3-labeled microarray assay have been described for HCV antibodies in addition to ELISA for the total HCV core antigen (Ag).
Results:
The recombinant immunoblot assay (RIBA) is a confirmatory test for HCV-Ab in blood, which is no longer needed due to the use of the sensitive third and fourth generation ELISA in addition to HCV-RNA detection by the Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). ELISA and RIBA measure current and prior exposure to HCV infection, but cannot discriminate between the two. RT-PCR is performed after ELISA for the diagnosis of HCV infection whether acute, chronic, false positive or false negative ELISA.
Conclusions:
The cooperation between ELISA and RT-PCR in the diagnosis of HCV infection has been tabulated and discussed. HCV genotyping and subtyping testing is essential in pre-treatment evaluation of the patients for setting valuable treatment strategies and in understanding the epidemiology of the virus.
