Qi Juju, Li Xinna, Zhang Yi, Shen Xinxin, Song Guowei, Pan Jing, Fan Tao, Wang Ruihuan, Li Lixin, Ma Xuejun
Hebei Medical University, 361 East Zhongshan Road, Shijiazhuang, 050031, Hebei, China.
Key Laboratory for Medical Virology, National Health and Family Planning Commission, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155 Changbai Road, Changping District, Beijing, 102206, China.
Arch Virol. 2019 Jul;164(7):1843-1850. doi: 10.1007/s00705-019-04230-z. Epub 2019 May 3.
Human respiratory syncytial virus (RSV) is a common viral pathogen that causes lower respiratory tract infections in infants and children globally. In this study, we developed a duplex reverse transcription recombinase-aided amplification (duplex-rtRAA) assay containing an internal control in a single closed tube for the detection of human RSV. The internal control in the amplification effectively eliminated false-negative results and ensured the accuracy of the duplex-rtRAA system. We first developed and evaluated a universal singleplex-rtRAA assay for RSV. The sensitivity of this assay for RSV was determined as 4.4 copies per reaction, and the specificity was 100%. Next, a duplex-rtRAA assay with an internal control was established. The sensitivity of the duplex-rtRAA assay approached 5.0 copies per reaction, and no cross-reaction with other common respiratory viruses was observed. The two detection methods (singleplex-rtRAA and duplex-rtRAA) developed in this study were used to test 278 clinical specimens, and the results showed absolute consistency with RSV RT-qPCR analysis, demonstrating 100% diagnostic sensitivity and specificity. These data indicate that the duplex-rtRAA has great potential for the rapid detection of RSV with a high sensitivity.
人呼吸道合胞病毒（RSV）是一种常见的病毒病原体，可在全球范围内导致婴幼儿下呼吸道感染。在本研究中，我们开发了一种在单个封闭管中包含内对照的双重逆转录重组酶辅助扩增（duplex-rtRAA）检测方法，用于检测人RSV。扩增过程中的内对照有效消除了假阴性结果，并确保了duplex-rtRAA系统的准确性。我们首先开发并评估了一种针对RSV的通用单重rtRAA检测方法。该检测方法对RSV的灵敏度确定为每个反应4.4个拷贝，特异性为100%。接下来，建立了一种带有内对照的duplex-rtRAA检测方法。duplex-rtRAA检测方法的灵敏度接近每个反应5.0个拷贝，未观察到与其他常见呼吸道病毒的交叉反应。本研究中开发的两种检测方法（单重rtRAA和duplex-rtRAA）用于检测278份临床标本，结果显示与RSV RT-qPCR分析完全一致，诊断灵敏度和特异性均为100%。这些数据表明，duplex-rtRAA在高灵敏度快速检测RSV方面具有巨大潜力。
