[Visual detection of HIV-1 by reverse transcription loop-mediated isothermal amplification with the hydroxynaphthol blue dye].

中华实验和临床病毒学杂志 Pub Date : 2013-04-01

Ya-Lan Zeng, Xiao-Guang Zhang, Kai Nei, Yi Zhang, Meng-Jie Yang, Hong-Wei Shen, Ji Wang, Lei Shi, Xue-Jun Ma

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引用次数: 0

Abstract

Objective: A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of HIV-1.

Methods: RT-LAMP primers were designed according to conservative sequences of HIV-1 gag gene, and their sensitivity and specificity were evaluated by the established RT-LAMP protocol with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The performance of RT-LAMP on clinical samples was compared with real-time reverse transcription PCR(qRT-PCR).

Results: The RT-LAMP assay showed a high specificity, and its detection limit was 1000 copies RNA per tube. The sensitivity and specificity of this method using 43 clinical samples were 94.6% and 100%, respectively,in comparison with those of qRT-PCR.

Conclusion: RT-LAMP assay using hydroxynaphthol blue dye does not need expensive instruments, and offer an alternative for the rapid detection of HIV-1 with the potential to be applied in field diagnosis.

本刊更多论文

[用羟基酚蓝染料逆转录环介导等温扩增目视检测HIV-1]。

目的:建立逆转录环介导的等温扩增(RT-LAMP)快速检测HIV-1的方法。方法:根据HIV-1 gag基因的保守序列设计RT-LAMP引物，在扩增前加入羟基酚蓝(HNB)染料，按照建立的RT-LAMP方案评价引物的敏感性和特异性。将RT-LAMP与实时反转录PCR(qRT-PCR)在临床样品上的表现进行比较。结果:RT-LAMP法特异性高，检出限为1000 copies RNA /管。与qRT-PCR相比，该方法对43份临床样本的敏感性和特异性分别为94.6%和100%。结论:使用羟基酚蓝染料的RT-LAMP检测方法不需要昂贵的仪器，为快速检测HIV-1提供了一种替代方法，具有应用于现场诊断的潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

中华实验和临床病毒学杂志

CiteScore

0.20

自引率

0.00%

发文量

4549