Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Kessler, HH; Santner, B; Umlauft, F; Urbanek, M; Kronawetter, M; Pierer, K; Stünzner, D; Grünewald, K; Marth, E.
Detection of hepatitis C viral sequences in serum by 'nested' polymerase chain reaction (PCR) and a commercial single-round PCR assay.
Clin Diagn Virol. 1995; 4(3):239-250
Doi: 10.1016/0928-0197(95)00014-Y
Web of Science
PubMed
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FullText_MUG
- Führende Autor*innen der Med Uni Graz
- Kessler Harald
- Co-Autor*innen der Med Uni Graz
- Marth Egon
- Santner Brigitte
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- Abstract:
- Background: Demonstration of the hepatitis C virus (HCV) genome is usually done with combined reverse transcription and polymerase chain reaction (RT-PCR) employing nested primer sets. Recently, a commercial PCR assay (Amplicor PCR assay), based on a simplified sample preparation procedure, a single, combined reverse transcription and polymerase chain reaction (RT-PCR), and a microwell plate capture and detection, has been developed. Objective: The aim of the present study was to compare the new Amplicor assay with an 'in-house' PCR. Additional testing included a third-generation enzyme immunoassay for anti-HCV antibodies, the Wellcozyme HCV Western Blot, which is equivalent to a third-generation recombinant immunoblot assay. Furthermore, HCV genotypes were classified. Study design: Sera from a total of 127 patients were studied. After screening with a third-generation enzyme immunoassay (EIA), the Wellcozyme HCV Western Blot, was performed as well as the conventional RT-PCR and the Amplicor PCR. Specimens, which were found positive by testing with the Amplicor kit, were subjected to storage at room temperature for 96 h. Results: A total of 52 patients were found to be positive for anti-HCV by the third-generation ELA. With the Amplicor assay, the HCV genome was detected in 38 patients. In comparison with the 'in-house' assay, two discrepant results were found. Resolution of discrepant samples increased the total number of true positives to 39. A good correlation was found between a positive anti-HCV test result and the presence of HCV-RNA by RT-PCR. No significant reduction in the amount of amplification product was observed by retesting of suboptimally stored samples with the Amplicor assay. Conclusion: Because of the rapidity and the improved ease of handling, the Amplicor assay was found to be a good contribution for detection of HCV in serum.
- Find related publications in this database (Keywords)
- Hepatitis C Virus
- Polymerase Chain Reaction
- Amplicor
- Detection
- Storage