F_0F_1-ATPase生物传感器快速检测食品中甲肝病毒

F_0F_1-ATPase Molecular Motor Biosensor for Rapid Detection of Hepatitis A Visus in Foods

建立一种应用F0F1-ATPase生物传感器快速检测甲肝病毒的方法。利用生物素-亲和素系统将甲肝病毒特异性分子探针连接在F0F1-ATPase的ε亚基上构建分子马达检测装置。将甲肝病毒阳性血清和阴性对照RNA分别与生物传感器结合,比较其启动ATP合成10min后的荧光强度的差别,对样品中甲肝病毒进行检测。结果表明,所建立的F0F1-ATPase生物传感器对检测食源性甲肝病毒具有良好的特异性和较高的灵敏度,对甲肝病毒核酸检测灵敏度达到0.01ng/mL,与反转录聚合酶链式反应检测结果作比较,结果具有良好一致性;本方法在1h内即可完成检测,可应用于食品中甲肝病毒的污染状况调查及高效检测。

An F0F1-ATPase molecular motor biosensor was established for rapid detection of Hepatitis A Virus （HAV） in foods. The biosensor was fabricated by connecting HIV specific probe to the ε subunit of F0F1-ATPase using an avidin- biotin system. The detection of HAV in samples was based on the difference in fluorescence intensity 10 min after initiation of ATP synthesis from HAV positive serum and negative control RNA when separately bound to the biosensor. The results demonstrated that the F0F1-ATPase biosensor exhibited good specificity and high sensitivity to HAV in foods. The sensitivity of the method was 0.01 ng/mL. The results obtained by this method agreed with those by reverse transcription PCR. The process of HAV detection with the biosensor can be completed within 1 h. In conclusion, this study has developed a method for the investigation and detection of HAV contamination in foods.