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通过逆转录环介导等温扩增技术快速检测尿液样本和感染的蚊子中的寨卡病毒。

Rapid Detection of Zika Virus in Urine Samples and Infected Mosquitos by Reverse Transcription-Loop-Mediated Isothermal Amplification.

机构信息

Department of Urology, Beaumont Health System, Royal Oak, MI, United States of America.

Oakland University William Beaumont School of Medicine, Rochester Hills, MI, United States of America.

出版信息

Sci Rep. 2018 Feb 28;8(1):3803. doi: 10.1038/s41598-018-22102-5.

Abstract

Infection with Zika virus (ZIKV) is of growing concern since infection is associated with the development of congenital neurological disease. Quantitative reverse transcription PCR (qRT-PCR) has been the standard for ZIKV detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper testing. Studies have suggested that ZIKV detection in urine is more sensitive and has a longer window of detection compared to serum and saliva. The objective of this study was to develop a urine diagnostic test that could be completed in under 30 minutes. Urine samples spiked with ZIKV or dengue virus were tested using RT-LAMP as well as by conventional quantitative qRT-PCR. These techniques were then validated using crude lysates made from ZIKV infected mosquitoes in addition to urine and serum samples from ZIKV infected patients. RT-LAMP specifically detected ZIKV in urine and serum for ZIKV infected patients and crude mosquito lysates. This test was performed in under 30 minutes and did not require RNA extraction from urine nor mosquitos. This approach could be used for monitoring of exposed individuals, especially pregnant women, couples wanting to conceive, or individuals with suspicious symptoms as well as surveillance of mosquito populations.

摘要

寨卡病毒(ZIKV)感染引起了越来越多的关注,因为感染与先天性神经疾病的发生有关。定量逆转录聚合酶链反应(qRT-PCR)一直是 ZIKV 检测的标准;然而,逆转录环介导等温扩增(RT-LAMP)可能允许更快和更便宜的测试。研究表明,与血清和唾液相比,尿液中 ZIKV 的检测更敏感,检测窗口更长。本研究的目的是开发一种可在 30 分钟内完成的尿液诊断测试。用 RT-LAMP 以及常规定量 qRT-PCR 检测用 ZIKV 或登革热病毒感染的尿液样本。然后,使用来自 ZIKV 感染的蚊子的粗提物以及来自 ZIKV 感染患者的尿液和血清样本对这些技术进行了验证。RT-LAMP 特异性地检测了 ZIKV 感染患者的尿液和血清中的 ZIKV 以及粗提物蚊子裂解物。该测试在 30 分钟内完成,不需要从尿液或蚊子中提取 RNA。这种方法可用于监测接触者,特别是孕妇、想要怀孕的夫妇或有可疑症状的个体,以及监测蚊子种群。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ddd/5830622/2fedcae32b80/41598_2018_22102_Fig1_HTML.jpg

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