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Updated: Jun 2, 2026

A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Diagnosis of hepatitis C virus infection by enzyme-linked immunosorbent assay and reverse transcriptase-nested
Menha Swellam1, Magda Sayed Mahmoud, Adel Abdel-Fatah Ali
1Department of Biochemistry, Genetic Engineering and Biotechnology Research Division, National Research Center, Dokki, Giza, Egypt. menha_m_swellam@hotmail.com
Abstract:
Hepatitis C virus is one of the main causes of chronic hepatitis in developing countries. The current study was to evaluate the efficacy of the enzyme-linked immunosorbent assay third generation (ELISA-3) for detection of antibodies to hepatitis C virus (anti-HCV) in comparison with reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) to detect HCV RNA for the diagnosis of hepatitis C virus. Serum samples were collected from 151 chronic hepatitis C patients and 50 healthy individuals. All samples were tested for anti-HCV antibodies using ELISA-3 and HCV RNA by RT-nested PCR. Of the 151,120 (78.9%) were found to be seropositive by ELISA-3, and 148 (98%) patients were HCV RNA positive, 118 (78.1%) were positives for both, 30 (19.9%) were positive for ELISA-3 and negative for RT-PCR, and 2 cases (1.3%) were positive for RT-nested PCR and negative for ELISA-3. The sensitivity and the specificity for the detection of HCV were absolute when the two techniques were combined. In conclusion, ELISA-3 is a suitable assay for routine screening for anti-HCV. RT-nested PCR for HCV is a value for the early detection of viremic, anti-HCV negative cases; this may be of importance in treatment of hepatitis C.
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Enzyme-Linked Immunosorbent Assay
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
