Xiao Xing-Long, Wu Hui, Li Yi-Juan, Li Hui-Fang, He Ya-Qing, Chen Gu, Zhang Jing-Wei, Yang Hong, Li Xiao-Feng, Yang Xiao-Quan, Yu Yi-Gang
Institution of Food safety, Department of Food, College of Light Industry & Food, South China University of Technology, Guangzhou 510640, China.
J Virol Methods. 2009 Jul;159(1):23-8. doi: 10.1016/j.jviromet.2009.02.022. Epub 2009 Mar 3.
Epidemics of acute hemorrhagic conjunctivitis are always explosive and extensive, and have been recognized as a serious international public health problem. Enterovirus 70 and coxsackievirus A24 variant have been identified as the major etiological agents in acute hemorrhagic conjunctivitis outbreaks worldwide. A novel multiplex real-time RT-PCR assay was developed for simultaneous detection, identification and quantitation of enterovirus 70 and coxsackievirus A24 variant. The specificity, sensitivity and reproducibility of the method were analyzed and 125 clinical samples were tested using this method. No cross-reactivity with other enteroviruses strains was detected. The detection limits achieved were 10 copies/tube of enterovirus 70 and 100 copies/tube of coxsackievirus A24 variant respectively, and the addition of the internal control does not compromise the sensitivity or specificity. One hundred and twenty five clinical samples were tested and the results were consistent with the results obtained by using virus isolation followed by neutralization and sequencing of VP1 region. Due to its high specificity, sensitivity and elimination of false negative results by the internal control, this assay is suitable for both research applications and rapid clinical diagnosis of enterovirus 70 and coxsackievirus A24 variant.
急性出血性结膜炎疫情总是呈爆发性且范围广泛，已被公认为一个严重的国际公共卫生问题。肠道病毒70型和柯萨奇病毒A24变种已被确定为全球急性出血性结膜炎疫情的主要病原体。开发了一种新型多重实时逆转录聚合酶链反应检测方法，用于同时检测、鉴定和定量肠道病毒70型和柯萨奇病毒A24变种。分析了该方法的特异性、敏感性和可重复性，并使用该方法检测了125份临床样本。未检测到与其他肠道病毒株的交叉反应。实现的检测限分别为肠道病毒70型每管10拷贝和柯萨奇病毒A24变种每管100拷贝，添加内部对照不会损害敏感性或特异性。检测了125份临床样本，结果与通过病毒分离后进行VP1区域中和及测序获得的结果一致。由于其高特异性、敏感性以及通过内部对照消除假阴性结果，该检测方法适用于肠道病毒70型和柯萨奇病毒A24变种的研究应用和快速临床诊断。
