Gao Mingxuan, Waggoner Jesse J, Hecht Sidney M, Chen Shengxi
Biodesign Center for BioEnergetics , Arizona State University , Tempe , Arizona 85287 , United States.
Department of Medicine, Division of Infectious Diseases , Emory University , Atlanta , Georgia 30322 , United States.
ACS Infect Dis. 2019 Nov 8;5(11):1907-1914. doi: 10.1021/acsinfecdis.9b00241. Epub 2019 Oct 1.
Dengue virus (DENV) is the most common human arboviral infection worldwide and can present with severe clinical manifestations. Timely DENV detection improves clinical outcomes, and identification of the DENV serotype (DENV-1-4) may provide beneficial epidemiologic data to inform the initiation of control measures. Here, DENV RNA-triggered, enzyme-free tandem toehold-mediated displacement reactions were developed to identify and serotype DENV in RNA controls and contrived samples through the amplification of a fluorescent signal detected by the use of a fluorescent scanner and a confocal microscope. Each DENV serotype was detected selectively using both imaging methods. In addition, a 384-well plate was used to prepare an array for diagnosis of the four DENV RNA serotypes from contrived clinical samples. The four serotypes of dengue virus were detected using novel enzyme-free amplification reactions, which are more facile than amplification using reverse transcriptase PCR.
登革病毒（DENV）是全球最常见的人类虫媒病毒感染，可表现出严重的临床症状。及时检测登革病毒可改善临床治疗效果，而鉴定登革病毒血清型（DENV-1-4）可为采取控制措施提供有益的流行病学数据。在此，我们开发了DENV RNA触发的、无酶串联趾hold介导的置换反应，通过使用荧光扫描仪和共聚焦显微镜检测到的荧光信号放大，来鉴定RNA对照和人工样本中的登革病毒并确定其血清型。使用这两种成像方法均可选择性地检测每种登革病毒血清型。此外，还使用384孔板制备了一个阵列，用于诊断人工临床样本中的四种登革病毒RNA血清型。使用新型无酶扩增反应检测了四种登革病毒血清型，该反应比使用逆转录酶PCR进行扩增更为简便。
