Visual detection of HIV-1 by reverse transcription loop-mediated isothermal amplification with the hydroxynaphthol blue dye
10.3760/cma.j.issn.1003-9279.2013.02.014
- VernacularTitle:基于颜色判定的逆转录环介导恒温扩增技术检测HIV-1病毒
- Keywords:
Nucleic acid amplification tecniques;
HIV-1;
Naphthols
- From:
Chinese Journal of Experimental and Clinical Virology
2013;27(2):126-128
- CountryChina
- Language:Chinese
-
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.
