Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis

Hofmann, Joerg; Frenzel, Katrin; Minh, Bui Q.; von Haeseler, Arndt; Edelmann, Anke; Ross, Stefan R.; Berg, Thomas; Krüger, Detlev H.; Meisel, Helga

doi:10.1016/j.diagmicrobio.2010.02.003

« Previous Next »Diagnostic Microbiology & Infectious Disease
Volume 67, Issue 2 , Pages 172-179, June 2010

Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis

Joerg Hofmann
- Institute of Medical Virology, Helmut-Ruska-Haus, Charité Universitätsmedizin, D-10117 Berlin, Germany
- Corresponding author. Institute of Medical Virology, Helmut-Ruska-Haus, Charité University Medicine Berlin, Charitéplatz 1, 10117 Berlin, Germany. Tel.: +49-30450525141; fax: +49-30450525141.
Katrin Frenzel
- Institute of Medical Virology, Helmut-Ruska-Haus, Charité Universitätsmedizin, D-10117 Berlin, Germany
Bui Q. Minh
- Center for Integrative Bioinformatics Vienna (CIBIV), Max F. Perutz Laboratories (MFPL), University of Vienna, Medical University of Vienna, University of Veterinary Medicine, A-1210 Vienna, Austria
Arndt von Haeseler
- Center for Integrative Bioinformatics Vienna (CIBIV), Max F. Perutz Laboratories (MFPL), University of Vienna, Medical University of Vienna, University of Veterinary Medicine, A-1210 Vienna, Austria
Anke Edelmann
- Institute of Medical Virology, Helmut-Ruska-Haus, Charité Universitätsmedizin, D-10117 Berlin, Germany
Stefan R. Ross
- Institute of Virology, Essen Universitätskrankenhaus, University of Duisburg-Essen, D-45141 Essen, Germany
Thomas Berg
- Klinik und Poliklinik für Gastroenterologie and Rheumatologie Universitätsklinikum Leipzig AöR, D-04006 Leipzog, Germany
Detlev H. Krüger
- Institute of Medical Virology, Helmut-Ruska-Haus, Charité Universitätsmedizin, D-10117 Berlin, Germany
Helga Meisel
- Institute of Medical Virology, Helmut-Ruska-Haus, Charité Universitätsmedizin, D-10117 Berlin, Germany

Received 2 November 2009; accepted 2 February 2010.

Abstract

Hepatitis D virus (HDV) infection is an important etiologic agent of fulminant hepatitis and may aggravate the clinical course of chronic hepatitis B infection resulting in cirrhosis and liver failure. This report describes the establishment of a real-time reverse transcriptase polymerase chain reaction method that allows the quantitative detection of HDV-1 and HDV-3 with a sensitivity in a linear range of 2 × 10³ to 10⁸ copies/mL. Additionally, the new assay provides the opportunity to distinguish HDV-1 from HDV-3 by a subsequent melting curve analysis, an important option because these HDV types are highly associated with severe clinical outcome. The results of the melting curve analysis of 42 HDV sequences obtained in this study and the phylogenetic analysis based on 139 full-length sequences from GenBank were consistent and showed that all sequences described here cluster within the HDV-1 clade. Therefore, this assay is useful for monitoring of antiviral treatment and molecular epidemiologic studies of HDV distribution.

Keywords: Hepatitis delta virus, Genotyping