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[丙型肝炎病毒基因分型不同方法的比较]

[Comparison of different methods of hepatitis C virus genotyping].

作者信息

Djebbi A, Sadraoui A, Triki H

机构信息

Laboratoire de virologie clinique, Institut Pasteur de Tunis, Tunisie.

出版信息

Ann Biol Clin (Paris). 2003 Nov-Dec;61(6):689-95.

Abstract

In order to introduce the approach of HCV genotyping in our laboratory, a comparative study of 3 molecular and 1 serological methods, was conducted on 62 HCV RNA positive sera. The molecular genotyping methods target the 5'untranslated (UTR) region of the virus genome and are based on an amplification of the viral genome, followed by partial sequencing, analyses of restriction fragment length polymorphisms (RFLP) or molecular hybridation (Inno LiPA, Innogenetics). The serological method or serotyping is based on the detection of antibodies to genotype specific epitopes derivated from the Non Structural (NS) 4 region of the viral genome (HCV 1-6 Serotyping Assay, Murex Biotech). "In house" methods, sequencing and RFLP, identified the genotype for 13 samples classified as non-typables by commercial kits Inno LiPA test and HCV 1-6 Serotyping Assay. Mixed infections revealed, especially by Inno LiPA, could not be identified by partial sequencing, which seems to detect only predominant genotype. For 4 samples, genotyping results of the methods targeting the 5'UTR were discordant with those of the serotyping of the NS4 region. Commercial kits are efficient to determine HCV genotypes, particularly in the context of antiviral therapy and patient's follow-up, sequencing remains the best alternative for more complete characterisation of viral strains and for epidemiological investigations.

摘要

为了介绍我们实验室丙型肝炎病毒(HCV)基因分型的方法,我们对62份HCV RNA阳性血清进行了3种分子方法和1种血清学方法的比较研究。分子基因分型方法针对病毒基因组的5'非翻译(UTR)区域,基于病毒基因组的扩增,随后进行部分测序、限制性片段长度多态性(RFLP)分析或分子杂交(Inno LiPA,Innogenetics)。血清学方法或血清分型基于检测源自病毒基因组非结构(NS)4区域的基因型特异性表位的抗体(HCV 1-6血清分型检测,Murex Biotech)。“内部”方法、测序和RFLP确定了13份样本的基因型,这些样本被商业试剂盒Inno LiPA检测和HCV 1-6血清分型检测分类为不可分型。混合感染尤其是通过Inno LiPA揭示的,无法通过部分测序鉴定,部分测序似乎只能检测到主要基因型。对于4份样本,针对5'UTR的方法的基因分型结果与NS4区域的血清分型结果不一致。商业试剂盒在确定HCV基因型方面是有效的,特别是在抗病毒治疗和患者随访的背景下,测序仍然是更全面地表征病毒株和进行流行病学调查的最佳选择。

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