Successful HCV genotyping of previously failed and low viral load specimens using an HCV RNA qualitative assay based on transcription-mediated amplification in conjunction with the line probe assay
Successful HCV genotyping of previously failed and low viral load specimens using an HCV RNA qualitative assay based on transcription-mediated amplification in conjunction with the line probe assay
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DOI：
10.1016/s1386-6532(02)00234-2
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发表时间：
2003-09-01
影响因子：
8.8
通讯作者：
Sherlock, C
中科院分区：
文献类型：
--
作者：
Comanor, L;Elkin, C;Sherlock, C
背景：丙型肝炎病毒（HCV）基因分型是慢性丙型肝炎诊断的关键部分。由Bayer Corporation销售的VERSANT（（R））HCV线性探针测定（利帕）需要PCR衍生的扩增子用于基因分型，所述扩增子通常从商业测定获得，包括Amplicor（（R））HCV 2.0（Amplicor 2.0）、Amplicor HCV Monitor（R））2.0或SuperQuant（（R））。偶尔，基于PCR的方法与利帕结合不能给出基因分型结果。尽管大多数基因分型失败发生在低病毒载量标本中，但也有一些发生在病毒载量相对较高的标本中。Bayer HCV RNA定性检测试剂盒（HCV TMA）的检测限约为5-10 IU/ml，比其他商业检测试剂盒更灵敏。目的：开发了使用HCV TMA与利帕连接（TMA-LiPA）的HCV基因分型方案，并测试了对先前通过基于PCR的方法与利帕联合进行基因分型失败的样本进行基因分型的能力。研究设计：临床样本从加拿大和美国的八个独立实验室获得，并在拜耳参考检测实验室使用TMA-LiPA进行检测。标本包括当基于PCR的检测与利帕联合使用时未能产生基因型结果的标本，以及因病毒载量低于原始实验室确定的经验证临界值而未尝试进行基因分型的标本。结果与结论：TMA-LiPA成功地对75份（90.7%）通过与利帕联合使用的基于PCR的方法进行基因分型失败的标本进行了基因分型，并对40份（90.0%）因病毒载量低而被拒绝进行基因分型的标本进行了基因分型。此外，TMA-LiPA为107例标本中的79例（73.8%）指定了亚型。我们的TMA-LiPA结果反映了在北美发现的HCV基因型的分布，并且与Amplicor 2.0联合利帕用于两种检测法基因分型的对照标本的结果100%一致。TMA-LiPA可能被证明在优化利帕性能和对患者标本进行基因分型方面都是有用的。(C)2002 Elsevier Science B. V.保留所有权利。
Background: Hepatitis C virus (HCV) genotyping is a critical part of the diagnostic work-up for chronic hepatitis C. The VERSANT((R)) HCV line probe assay (LiPA) marketed by Bayer Corporation requires PCR-derived amplicons for genotyping usually obtained from commercial assays, including Amplicor((R)) HCV 2.0 (Amplicor 2.0), Amplicor HCV Monitor((R)) 2.0, or SuperQuant((R)). Occasionally, PCR-based methods in conjunction with LiPA fail to give a genotyping result. Although most genotyping failures occur among low viral load specimens, some occur in specimens with relatively high viral loads. The Bayer HCV RNA Qualitative assay (HCV TMA), with a limit of detection of approximately 5-10 IU/ml, is more sensitive than other commercial assays. Objectives: An HCV genotyping protocol using HCV TMA linked with LiPA (TMA-LiPA) was developed and tested for ability to genotype samples that had previously failed genotyping by PCR-based methods in conjunction with LiPA. Study design: Clinical specimens were obtained from eight independent laboratories in Canada and the US and tested with TMA-LiPA at the Bayer Reference Testing Laboratory. Specimens included those that failed to produce a genotype result when a PCR-based assay was used in conjunction with LiPA and specimens for which genotyping was not attempted because the viral load was below the validated cut-off determined in the laboratory of origin. Results and conclusions: TMA-LiPA successfully genotyped 68 of 75 (90.7%) specimens that had failed genotyping by PCR-based methods used in conjunction with LiPA and 36 of 40 (90.0'Yo) specimens that were rejected for genotyping due to low viral load. Moreover, TMA-LiPA assigned subtype for 79 of 107 (73.8%) specimens. Our TMA-LiPA results reflected the distribution of HCV genotypes found in North America, and were 100% concordant with those of Amplicor 2.0 in conjunction with LiPA for control specimens genotyped by both assays. TMA-LiPA may prove useful both in optimizing LiPA performance and genotyping patient specimens. (C) 2002 Elsevier Science B.V. All rights reserved.