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

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

doi:10.1016/j.diagmicrobio.2010.02.003

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

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

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 × 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.

Original language	English
Pages (from-to)	172-179
Number of pages	8
Journal	Diagnostic Microbiology and Infectious Disease
Volume	67
Issue number	2
DOIs	https://doi.org/10.1016/j.diagmicrobio.2010.02.003
Publication status	Published - Jun 2010
Externally published	Yes

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Hofmann, J., Frenzel, K., Minh, B. Q., von Haeseler, A., Edelmann, A., Ross, S. R., Berg, T., Krüger, D. H., & Meisel, H. (2010). Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis. Diagnostic Microbiology and Infectious Disease, 67(2), 172-179. https://doi.org/10.1016/j.diagmicrobio.2010.02.003

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

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 × 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.",

keywords = "Genotyping, Hepatitis delta virus",

author = "Joerg Hofmann and Katrin Frenzel and Minh, {Bui Q.} and {von Haeseler}, Arndt and Anke Edelmann and Ross, {Stefan R.} and Thomas Berg and Kr{\"u}ger, {Detlev H.} and Helga Meisel",

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language = "English",

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Hofmann, J, Frenzel, K, Minh, BQ, von Haeseler, A, Edelmann, A, Ross, SR, Berg, T, Krüger, DH & Meisel, H 2010, 'Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis', Diagnostic Microbiology and Infectious Disease, vol. 67, no. 2, pp. 172-179. https://doi.org/10.1016/j.diagmicrobio.2010.02.003

TY - JOUR

T1 - Quantitative detection and typing of hepatitis D virus in human serum by real-time polymerase chain reaction and melting curve analysis

AU - Hofmann, Joerg

AU - Frenzel, Katrin

AU - Minh, Bui Q.

AU - von Haeseler, Arndt

AU - Edelmann, Anke

AU - Ross, Stefan R.

AU - Berg, Thomas

AU - Krüger, Detlev H.

AU - Meisel, Helga

PY - 2010/6

Y1 - 2010/6

N2 - 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 × 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.

AB - 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 × 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.

KW - Genotyping

KW - Hepatitis delta virus

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U2 - 10.1016/j.diagmicrobio.2010.02.003

DO - 10.1016/j.diagmicrobio.2010.02.003

M3 - Article

SN - 0732-8893

VL - 67

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EP - 179

JO - Diagnostic Microbiology and Infectious Disease

JF - Diagnostic Microbiology and Infectious Disease

IS - 2

ER -

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

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