PDF(Pubmed)
Abstract：
Enteroviruses (EVs), which belong to the Picornaviridae family, infect individuals asymptomatically or cause mild symptoms (fever, runny nose, cough, skin rash, sneezing, mouth blister). Severe cases can cause various diseases, such as acute hemorrhagic conjunctivitis, aseptic meningitis, or myocarditis, especially in infants. These viruses can be transmitted via the fecal-oral route via contaminated water. In this study, we established a polymerase chain reaction (PCR) method for detecting EVs in water sample using Coxsackievirus B5 (CV-B5) and Echovirus 30 (E-30), which belong to species B of the four species of EVs (EV-A to D). Several methods have been investigated and compared for the detection of EVs, including real-time reverse transcription (RT) polymerase chain reaction and conventional RT-PCR. The most sensitive primer sets were selected, and the PCR conditions were modified to increase sensitivity. We also quantified the detection limits of real-time and conventional RT-PCR. The detection limits of conventional RT-PCR were detected in 105-106 copy/mL for CV-B5 and 106-107 copy/mL for E-30, respectively. This optimized method for detecting EVs is expected to contribute substantially to the investigation of EV outbreaks in water samples.
摘要：
肠道病毒（EV），属于小导航科，无症状地感染个体或引起轻度症状(发烧，流鼻涕,咳嗽,皮疹,打喷嚏,嘴巴起泡)。严重的病例会导致各种疾病，如急性出血性结膜炎，无菌性脑膜炎,或者心肌炎,尤其是婴儿。这些病毒可以通过粪便-口腔途径通过污染的水传播。在这项研究中,我们建立了使用柯萨奇病毒B5（CV-B5）和Echovirus30（E-30）检测水样中EV的聚合酶链反应（PCR）方法，属于四种电动汽车(EV-A至D)中的B种。已经研究并比较了几种检测电动汽车的方法，包括实时逆转录(RT)聚合酶链反应和常规RT-PCR。选择最敏感的引物组，并修改PCR条件以增加灵敏度。我们还定量了实时和常规RT-PCR的检测限。常规RT-PCR的检测限分别为CV-B5的105-106拷贝/mL和E-30的106-107拷贝/mL。这种用于检测EV的优化方法预计将大大有助于水样中EV爆发的调查。
