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通过分子马达生物传感器技术建立一种特异、便捷、快速的食源性轮状病毒检测方法。以F0F1-ATPase为核心构建分子马达,以轮状病毒保守片段VP7设计各血清型通用探针,通过生物素-亲和素系统将探针与分子马达连接构建F0F1-ATPase分子马达检测装置。提取病毒RNA并将其与生物传感器结合的同时启动ATP合成,比较其荧光强度的差别,可以对样品中的RNA进行检测。此方法的病毒RNA检测灵敏度为0.005 ng/mL,对轮状病毒检测特异,与甲肝病毒、诺如病毒无交叉反应,在1 h内即可完成检测。运用此方法随机检测15份样品,检测结果与RT-PCR一致。结果表明,分子马达生物传感器检测轮状病毒的方法灵敏、特异,可用于食源性轮状病毒的快速检测。
Through the molecular motor biosensor technology to establish a specific, convenient and rapid detection of foodborne rotavirus. A molecular motor was constructed with F0F1-ATPase as the core. The universal probe of serotypes was designed with the rotavirus conserved fragment VP7. The probe was connected with the molecular motor via biotin-avidin system to construct F0F1-ATPase molecular motor detection device. Extraction of viral RNA and its combination with the biosensor to start ATP synthesis, comparing the differences in fluorescence intensity, RNA samples can be detected. The detection sensitivity of the virus RNA of this method is 0.005 ng / mL, which is specific to rotavirus and has no cross-reaction with Hepatitis A and Norovirus and can be detected within 1 h. Using this method, 15 samples were randomly detected and the results were consistent with RT-PCR. The results showed that the molecular motor biosensor detection of rotavirus sensitive and specific, can be used for rapid detection of foodborne rotavirus.