CAJ | 학술논문

目的建立一种基于颜色判定的用于人类免疫缺陷病毒Ⅰ型(HIV-1)检测的逆转录环介导恒温扩增(RT-LAMP)技术.方法根据HIV-1 gag基因保守区的序列设计RT-LAMP引物,利用已建立好的基于羟基萘酚蓝(HNB)颜色判定的RT-LAMP体系验证其灵敏度与特异性,并对实时荧光逆转录PCR(qRT-PCR)确认的临床样本进行一致性对比.结果 RT-LAMP引物特异性高,检出限为1000个拷贝RNA,对43份临床样本检测的灵敏度和特异性分别为94.6％～ 100％.结论基于颜色判定的环介导恒温扩增方法摆脱了对昂贵仪器的依赖,有望应用于HIV-1感染的现场筛选,为快速检测HIV-1病毒提供了新方法和新思路.
목적 건립일충기우안색판정적용우인류면역결함병독Ⅰ형(HIV-1)검측적역전록배개도항온확증(RT-LAMP)기술.방법 근거HIV-1 gag기인보수구적서렬설계RT-LAMP인물,이용 이건립호적기우간기내분람(HNB)안색판정적RT-LAMP체계험증기령민도여특이성,병대실시형광역전록PCR(qRT-PCR)학인적림상양본진행일치성대비.결과 RT-LAMP인물특이성고,검출한위1000개고패RNA,대43빈림상양본검측적령민도화특이성분별위94.6％～ 100％.결론 기우안색판정적배개도항온확증방법파탈료대앙귀의기적의뢰,유망응용우HIV-1감염적현장사선,위쾌속검측HIV-1병독제공료신방법화신사로.
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.