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用于检测HIV-1耐药性突变的LiPA HIV-1 RT检测、选择性PCR和直接固相测序的比较。

Comparison of the LiPA HIV-1 RT test, selective PCR and direct solid phase sequencing for the detection of HIV-1 drug resistance mutations.

作者信息

Schmit J C, Ruiz L, Stuyver L, Van Laethem K, Vanderlinden I, Puig T, Rossau R, Desmyter J, De Clercq E, Clotet B, Vandamme A M

机构信息

Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium.

出版信息

J Virol Methods. 1998 Jul;73(1):77-82. doi: 10.1016/s0166-0934(98)00043-3.

Abstract

The performance to detect drug resistance mutations in the reverse transcriptase gene of HIV-1 was compared for direct solid phase sequencing, selective polymerase chain reaction (PCR) using the amplification refractory mutation system (ARMS) and the new line probe assay (LIPA) HIV-1 RT. The three tests were undertaken on 50 plasma samples from 25 treatment-experienced patients under combination therapy with dideoxynucleoside analogues. LiPA HIV-1 RT gave interpretable results in 80 to 96% of the samples depending on the codon of interest. In 2% of the samples a failure to amplify resulted in uninterpretable results for sequencing. ARMS gave no result in seven samples (14%). Overall, there was a 73 to 100% concordance between the three methods. In this study, LiPA HIV-1 RT proved to be an accurate and reliable alternative to DNA sequencing for the detection of drug resistance mutations in patient samples.

摘要

对直接固相测序、使用扩增阻滞突变系统(ARMS)的选择性聚合酶链反应(PCR)以及新型线性探针分析(LIPA)HIV-1逆转录酶检测HIV-1逆转录酶基因耐药性突变的性能进行了比较。对25例接受双脱氧核苷类似物联合治疗的有治疗经验患者的50份血浆样本进行了这三项检测。根据所关注的密码子,LiPA HIV-1 RT在80%至96%的样本中给出了可解释的结果。在2%的样本中,扩增失败导致测序结果无法解释。ARMS在7个样本(14%)中未得出结果。总体而言,三种方法之间的一致性为73%至100%。在本研究中,LiPA HIV-1 RT被证明是一种准确可靠的替代DNA测序的方法,用于检测患者样本中的耐药性突变。

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