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开发一种便携式逆转录环介导等温扩增系统,以经济有效的方式检测基孔肯雅病毒的 E1 区域。

Development of a portable reverse transcription loop-mediated isothermal amplification system to detect the E1 region of Chikungunya virus in a cost-effective manner.

机构信息

Department of Infectious Disease Control, Faculty of Medicine, Oita University, Oita, Japan.

Nutrition and Clinical Services Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr, b), Dhaka, Bangladesh.

出版信息

Genes Cells. 2020 Sep;25(9):615-625. doi: 10.1111/gtc.12797. Epub 2020 Jul 22.

Abstract

Chikungunya fever is a mosquito-borne disease cause of persistent arthralgia. The current diagnosis of Chikungunya virus (CHIKV) relies on a conventional reverse transcription polymerase chain reaction assay. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a rapid and simple tool used for DNA-based diagnosis of a variety of infectious diseases. In this study, we established an RT-LAMP system to recognize CHIKV by targeting the envelope protein 1 (E1) gene that could also detect CHIKV at a concentration of 8 PFU without incorrectly detecting other mosquito-borne viruses. The system also amplified the E1 genome in the serum of CHIKV-infected mice with high sensitivity and specificity. Moreover, we established a dry RT-LAMP system that can be transported without a cold chain, which detected the virus genome in CHIKV-infected patient samples with high accuracy. Thus, the dry RT-LAMP system has great potential to be applied as a novel CHIKV screening kit in endemic areas.

摘要

基孔肯雅热是一种由蚊子传播的疾病,会导致持续性关节痛。目前基孔肯雅病毒(CHIKV)的诊断依赖于传统的反转录聚合酶链反应检测。反转录环介导等温扩增(RT-LAMP)是一种快速而简单的工具,用于基于 DNA 的各种传染病的诊断。在本研究中,我们建立了一种 RT-LAMP 系统,通过针对包膜蛋白 1(E1)基因来识别 CHIKV,该系统可以在 8PFU 的浓度下检测到 CHIKV,而不会错误地检测到其他蚊媒病毒。该系统还可以高度敏感和特异性地扩增感染 CHIKV 的小鼠血清中的 E1 基因组。此外,我们建立了一种干燥的 RT-LAMP 系统,无需冷链运输,可高度准确地检测到感染 CHIKV 的患者样本中的病毒基因组。因此,干燥的 RT-LAMP 系统具有作为流行地区新型 CHIKV 筛查试剂盒的巨大应用潜力。

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