Visual detection of HIV-1 by reverse transcription loop-mediated isothermal amplification with the hydroxynaphthol blue dye.
- Author:
Ya-Lan ZENG
;
Xiao-Guang ZHANG
;
Kai NEI
;
Yi ZHANG
;
Meng-Jie YANG
;
Hong-Wei SHEN
;
Ji WANG
;
Lei SHI
;
Xue-Jun MA
- Publication Type:Journal Article
- MeSH: HIV-1; isolation & purification; Naphthalenesulfonates; Nucleic Acid Amplification Techniques; methods; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; Sensitivity and Specificity
- From: Chinese Journal of Experimental and Clinical Virology 2013;27(2):126-128
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEA reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of HIV-1.
METHODSRT-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).
RESULTSThe 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.
CONCLUSIONRT-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.