Evaluation and comparison of different hepatitis C virus genotyping and serotyping assays

• Published in: Journal of hepatology Vol. 26; no. 5; pp. 1001 - 1009
• Main Authors: Lee, Jung-Hun, Roth, W.Kurt, Zeuzem, Stefan
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier B.V 01.05.1997; Elsevier
• Subjects: Biological and medical sciences; DNA, Viral - genetics; Evaluation Studies as Topic; Female; Genotype; Genotyping assays; Hepacivirus - classification; Hepacivirus - genetics; Hepatitis C virus genotype/subtype; Human viral diseases; Humans; Immunoassay; Immunoblotting; Immunoenzyme Techniques; Infectious diseases; Male; Medical sciences; Nucleic Acid Hybridization; Phylogenetic analysis; Polymerase chain reactioni; Serotyping; Serotyping assays; Viral diseases; Viral hepatitis; Phylogenetic analysis; Genotyping assays; Hepatitis C virus genotype/subtype; Polymerase chain reactioni; Serotyping assays; Human; Immunopathology; Evaluation; Genetic inheritance; Hepatic disease; Phylogeny; Biological activity; Infection; Polymerase chain reaction; Chemotherapy; Serotype specificity; Viral disease; Digestive diseases; Interferon; Predictive factor; Comparative study; Viral hepatitis C
• ISSN: 0168-8278; 1600-0641
• DOI: 10.1016/S0168-8278(97)80108-0

Background/Aims: Evidence that the geno/subtype of hepatitis C virus (HCV) is predictive of the response to initerferon-α therapy suggests that typing methods are clinical useful. In the present study, HCV isolates obtained from 74 patients with chronic hepatitis C were used to evaluate three genotyping and two serotyping assays. Methods: The reverse hybridization assay and the DNA immunoassay are based on immobilized type-specific probes for the 5′-noncoding and the core region, respectively. A third genotyping assay utilized type-specific primers for amplification of the core region. Serotyping assays detect type-specific antibodies of the nonstructural-4 region (enzyme immunoassay) or of the core and nonstructural-4 region (recombinant immunoblot assay). Gold standard geno/subtyping of HCV isolates was performed by sequency and phylogenetic analysis of the nonstructural-5B region. Results: All genotyping systems amplified the respective target region of the HCV genome with high sensitivity. The reverse hybridization assay and the DNA immunoassay correctly identified HCV-1, -2, and -3. The DNA immunoassay misinterpreted all HCV-4 isolates as HCV-4 and -5 coinfection. In the type-specific amplification assay, coinfections of subtypes HCV-1a and HCV-3a with HCV-1b could not be excluded. The reverse hybridization assay misinterpreted 1/14 HCV-1a isolates as HCV-1b, and vice versa 3/36 HCV-1b isolates as HCV-1a. Furthermore, differentiation between HCV-2a and -2c was not possible using this assay. The DNA immunoassay correctly identified all HCV subtypes. The serotyping assays, recombinant immunoblot assay and enzyme immunoassay identified HCV-1, -2, and -3 in 93% and 89% of cases, respectively. HCV-4, however, could only be recognized by the enzyme immunoassay. Conclusions: The reverse hybridization assay and the DNA immunoassay specifically identified HCV genotypes 1, 2, and 3, while crossreactivity occurred in the primer-specific amplification assay. The DNA immunoassay achieved the best performance in HCV subtyping. Both serotyping systems correctly identified HCV-1, -2, and -3 in about 90% of cases, but lack the possibility of subtyping.