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Eur J Clin Microbiol Infect Dis. 2008 Dec;27(12):1177-82. doi: 10.1007/s10096-008-0556-9. Epub 2008 Jun 13.

The development of a qualitative real-time RT-PCR assay for the detection of hepatitis C virus.

Author information

1
Department of Clinical Microbiology, St. James's Hospital, Dublin 8, Ireland. aclancy@stjames.ie

Abstract

Real-time polymerase chain reaction (PCR) represents a favourable option for the detection of hepatitis C virus (HCV). A real-time reverse transcriptase PCR (RT-PCR) assay was developed as a qualitative diagnostic screening method for the detection of HCV using the ABI PRISM 7500 Sequence Detection System. The primers and probe were designed to target the 5'-untranslated region of the hepatitis C viral genome. A second heterologous probe assay was developed for the detection of the haemagglutinin gene of phocine distemper virus (PDV) and was used as an internal control. A semi-automated HCV extraction method was also implemented using the ABI PRISM 6100 Nucleic Acid PrepStation. The HCV assay was optimised as a qualitative singleplex RT-PCR assay with parallel testing of the target and internal control. The assay results (n = 200) were compared to the COBAS AMPLICOR HCV Test v2.0 assay. The assay demonstrated a high rate of sensitivity (99%), specificity (100%) and an acceptable limit of detection (LOD) of 100 IU/ml. The development of a qualitative multiplex assay for the simultaneous detection of HCV and internal control indicates the same high rates of sensitivity and specificity. This sensitive real-time assay may prove to be a valuable method for the detection of HCV.

PMID:
18551325
DOI:
10.1007/s10096-008-0556-9
[Indexed for MEDLINE]

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