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用于检测HIV-1耐药性的线性探针分析(LIPA)与序列分析的比较。

Comparison of line probe assay (LIPA) and sequence analysis for detection of HIV-1 drug resistance.

作者信息

Puchhammer-Stöckl E, Schmied B, Mandl C W, Vetter N, Heinz F X

机构信息

Institute of Virology, University of Vienna, Austria.

出版信息

J Med Virol. 1999 Mar;57(3):283-9. doi: 10.1002/(sici)1096-9071(199903)57:3<283::aid-jmv12>3.0.co;2-2.

Abstract

The identification of HIV strains that are resistant to antiretroviral drugs, which emerge during a patient's therapy or are already present in infected individuals prior to treatment, is of increasing importance for the clinical management of HIV-infected persons. Two different methods were compared for the genotypic detection of resistance development in the HIV-1 reverse transcriptase (RT) gene, the commonly used sequence analysis, and the commercially available RT-line immunoprobe assay (LIPA), which can detect mutations at six separate codons of the RT gene, which are known to confer resistance to certain nucleoside inhibitors. Eighty serum samples from HIV-1-infected persons, some of whom were receiving antiretroviral therapy, were investigated in parallel by sequencing as well as by LIPA. LIPA results agreed with sequence data in the vast majority of the cases. However, in 40% of the samples, LIPA failed to yield evaluable results for one or more of the codon positions. In particular, LIPA detection rate was low at codon 41 (75%), whereas at codons 69/70, 74, 184, and 215 results were obtained from 90%-95% of the samples. A number of mutations in the vicinity of the respective codons were detected by sequencing, and these may have been responsible for the LIPA hybridization failure. There remained a number of samples, however, where no explanation for the lack of hybridization could be derived from sequence data. Our results indicate that the use of the LIPA does not eliminate the need for sequence analysis for detection of drug-resistant HIV strains.

摘要

在患者治疗过程中出现的或在治疗前已存在于感染者体内的对抗逆转录病毒药物耐药的HIV毒株的鉴定,对于HIV感染者的临床管理愈发重要。对两种不同方法进行了比较,以用于HIV-1逆转录酶(RT)基因耐药性发展的基因分型检测,一种是常用的序列分析,另一种是市售的RT-线性免疫探针分析(LIPA),后者可检测RT基因六个不同密码子处的突变,已知这些突变会导致对某些核苷抑制剂产生耐药性。对80份来自HIV-1感染者的血清样本进行了研究,其中一些人正在接受抗逆转录病毒治疗,同时采用测序和LIPA方法进行检测。在绝大多数情况下,LIPA结果与序列数据一致。然而,在40%的样本中,LIPA未能得出一个或多个密码子位置的可评估结果。特别是,密码子41处的LIPA检测率较低(75%),而在密码子69/70、74、184和215处,90%-95%的样本获得了结果。通过测序在各个密码子附近检测到了一些突变,这些突变可能是导致LIPA杂交失败的原因。然而,仍有一些样本,从序列数据中无法找到杂交失败的原因。我们的结果表明,使用LIPA并不能消除对耐药HIV毒株检测进行序列分析的必要性。

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