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Visual detection of Zika virus by isothermal nucleic acid amplification combined with a lateral-flow device

机译:通过等温核酸扩增结合侧向流动装置目视检测寨卡病毒

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The Zika virus (ZIKV) did not receive significant attention in the past until the ZIKV outbreak occurred a few years ago. It has been shown that ZIKV can trigger congenital microcephaly, Guillain–Barré syndrome and other neurological syndromes. To fight against ZIKV, the efficient diagnosis of ZIKV is absolutely required; this has prompted us to establish a visual detection method for ZIKV with high accuracy and sensitivity. We applied reverse transcription loop-mediated isothermal amplification (RT-LAMP) with primers targeted to the specific conserved region of the non-structural protein 5 (NS5) gene fragment; moreover, using a lateral flow device (LFD), the detection of the ZIKA genome was completed within 1 hour in a 65 °C water bath. Compared with one-step real-time PCR (one-step RT-PCR), a RT-LAMP-turbidimeter, and quantitative reverse transcription PCR (RT-qPCR), our method is more convenient, sensitive, and specific, less time-consuming, and has equal detection performance. The newly developed method was evaluated for 12 clinical serum samples, and the results were consistent with the previous RT-qPCR detection results obtained by the Centers for Disease Control and Prevention of Guangdong; this supported that the developed method could be a potential solution for ZIKV diagnosis.
机译:直到几年前ZIKV爆发之前,寨卡病毒(ZIKV)才受到关注。研究表明,ZIKV可以引发先天性小头畸形,格林-巴利综合征和其他神经系统综合征。为了对抗ZIKV,绝对需要对ZIKV进行有效的诊断。这促使我们建立了具有高精度和高灵敏度的ZIKV视觉检测方法。我们应用了针对非结构蛋白5(NS5)基因片段的特定保守区域的引物进行逆转录环介导的等温扩增(RT-LAMP);此外,使用侧向流动装置(LFD),在65°C水浴中在1小时内完成了ZIKA基因组的检测。与一步式实时PCR(一步式RT-PCR),RT-LAMP-浊度计和定量逆转录PCR(RT-qPCR)相比,我们的方法更方便,灵敏,特异,且所需时间更少耗电量大,并且具有相同的检测性能。对新开发的方法对12种临床血清样品进行了评估,结果与广东省疾病预防控制中心以前的RT-qPCR检测结果一致;这支持开发的方法可能是ZIKV诊断的潜在解决方案。

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