Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis.
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 x 10(3) to 10(8) 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.
Hofmann
Joerg
J
Institute of Medical Virology, Helmut-Ruska-Haus, Charité Universitätsmedizin, D-10117 Berlin, Germany. joerg.hofmann@charite.de
Frenzel
Katrin
K
Minh
Bui Q
BQ
von Haeseler
Arndt
A
Edelmann
Anke
A
Ross
Stefan R
SR
Berg
Thomas
T
Krüger
Detlev H
DH
Meisel
Helga
H
D016000:Cluster Analysis; D003699:Hepatitis D / Q000175:diagnosis* / Q000821:virology; D003698:Hepatitis Delta Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D044967:Serum / Q000821:virology*; D044366:Transition Temperature; D014773:Virology / Q000379:methods*
D012367:RNA, Viral