Ross R S, Viazov S, Kpakiwa S S, Roggendorf M
Institute of Virology, National Reference Centre for Hepatitis C, Essen University Hospital, University of Duisburg-Essen, Essen, Germany.
J Clin Lab Anal. 2007;21(5):340-7. doi: 10.1002/jcla.20195.
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.
丙型肝炎病毒（HCV）分离株的分型目前是合理定制抗病毒联合治疗的前提条件。在许多诊断实验室中，似乎有一种倾向，即采用利用扩增产物的便捷省时程序，这些扩增产物可从先前的定性或定量HCV核糖核酸（RNA）检测中获得。在此背景下，我们评估了一种技术组合的性能特征，即转录介导扩增-线性探针分析（TMA-LiPA），它将HCV RNA的高灵敏度TMA与VERSANT HCV基因型分析（第1版）相联系。总共100份临床样本通过TMA-LiPA进行基因分型。将获得的结果与基于原始巢式逆转录（RT）-聚合酶链反应（PCR）的VERSANT检测、核心相关的GEN-ETI-K DEIA以及HCV核心和NS5B区域部分序列的系统发育分析所记录的结果进行比较。TMA-LiPA为所有100株HCV分离株正确分型。对于1型和2型分离株的亚型分型，TMA-LiPA的鉴别能力分别仅为82%和53%。因此，在我们手中，TMA-LiPA成为一种便捷省时的HCV分型常规程序，目前可为临床目的提供足够的信息。然而，与所有基于5'非翻译区（UTR）的检测一样，该技术在解析现有HCV亚型复杂性的潜力方面存在局限性。
