Open AccessTranscription‐mediated amplification linked to line probe assay as a routine tool for HCV typing in clinical laboratories
Author(s) -
Ross R.S.,
Viazov S.,
Kpakiwa S.S.,
Roggendorf M.
Publication year - 2007
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.20195
Subject(s) - subtyping , ns5b , typing , virology , genotype , hepatitis c virus , biology , genotyping , amplicon , polymerase chain reaction , microbiology and biotechnology , computational biology , genetics , virus , hepacivirus , gene , computer science , programming language
Typing of hepatitis C virus (HCV) isolates is currently a prerequisite for adequate tailoring of antiviral combination therapy. In many diagnostic laboratories, there seems to be a tendency toward convenient and time‐saving procedures utilizing amplification products, which are already available from preceding qualitative or quantitative HCV ribonucleic acid (RNA) assays. In this context, we evaluated the performance characteristics of a combination of techniques, i.e., transcription‐mediated amplification‐line probe assay (TMA‐LiPA), which links highly sensitive TMA of HCV RNA to the VERSANT HCV Genotype Assay (version 1). A total of 100 clinical samples were genotyped by TMA‐LiPA. The obtained results were compared to those recorded by the original, nested reverse transcription (RT)‐polymerase chain reaction (PCR)‐based VERSANT assay, the core‐related GEN‐ETI‐K DEIA, and phylogenetic analyses of partial sequences from the HCV core and NS5B regions. TMA‐LiPA assigned the correct genotype to all 100 HCV isolates. For subtyping of genotype 1 and 2 isolates, TMA‐LiPA only showed discriminatory powers of 82% and 53%, respectively. Thus, TMA‐LiPA in our hands turned out as a convenient and time‐saving routine procedure for HCV typing which currently provides sufficient information for clinical purposes. Like all 5′untranslated region (UTR)‐based assays, the technique is limited, however, in its potentials to resolve the complexity of existing HCV subtypes. J. Clin. Lab. Anal. 21:340–347, 2007. © 2007 Wiley‐Liss, Inc.