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Utilization of recombinase polymerase amplification method combined with lateral flow dipstick for visual detection of respiratory syncytial virus.
Molecular and Cellular Probes ( IF 3.3 ) Pub Date : 2019-10-22 , DOI: 10.1016/j.mcp.2019.101473
Yu-Zhong Xu 1 , Du-Zhi Fang 2 , Fang-Fang Chen 3 , Qin-Fei Zhao 3 , Chao-Ming Cai 2 , Ming-Gang Cheng 2
Affiliation  

Respiratory syncytial virus (RSV) is a major causative agent of respiratory tract infection necessitating hospitalization in children. A rapid diagnostic method would facilitate early detection of RSV infection and timely implementation of special treatment. Here, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with lateral flow dipstick (LFD) was evaluated for rapid visual detection of RSV. The primers were designed to target the conserved L gene. The RT-RPA-LFD assay could simultaneously detect RSV subtype A and B with the same detection limit of 10 copies of a given RNA molecule. Moreover, the assay showed no cross-reactivity with other common human pathogens. The performance of the RT-RPA-LFD assay was evaluated by testing 136 nasopharyngeal aspirates (NPAs). The agreement of the detection results between RT-RPA-LFD and qRT-PCR was 100% (34 positive and 102 negative cases). In summary, the developed RT-RPA-LFD assay had good performance in detecting RSV in clinical specimens, thus providing a novel alternative solution for the detection of RSV under field conditions.

中文翻译:

利用重组酶聚合酶扩增法结合侧向量油尺对呼吸道合胞病毒进行视觉检测。

呼吸道合胞病毒(RSV)是呼吸道感染的主要病原体,需要儿童住院治疗。快速诊断方法将有助于及早发现RSV感染并及时实施特殊治疗。在这里,逆转录重组酶聚合酶扩增(RT-RPA)分析结合侧向量油尺(LFD)进行了评估,以快速目测RSV。设计引物以靶向保守的L基因。RT-RPA-LFD检测可以同时检测RSV亚型A和B,而给定RNA分子的10个拷贝的检测限相同。而且,该测定显示与其他常见人类病原体没有交叉反应。通过测试136鼻咽抽吸物(NPA)评估了RT-RPA-LFD分析的性能。RT-RPA-LFD和qRT-PCR检测结果的一致性为100%(34例阳性和102例阴性)。综上所述,开发的RT-RPA-LFD检测方法在临床标本中检测RSV具有良好的性能,从而为野外条件下RSV的检测提供了新的替代解决方案。
更新日期:2019-11-04
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