1.Visual detection of HIV-1 by reverse transcription loop-mediated isothermal amplification with the hydroxynaphthol blue dye.
Ya-Lan ZENG ; Xiao-Guang ZHANG ; Kai NEI ; Yi ZHANG ; Meng-Jie YANG ; Hong-Wei SHEN ; Ji WANG ; Lei SHI ; Xue-Jun MA
Chinese Journal of Experimental and Clinical Virology 2013;27(2):126-128
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.
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
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