食品研究与开发
食品研究與開髮
식품연구여개발
FOOD RESEARCH AND CEVELOPMENT
2014年
24期
116-120
,共5页
张捷%许美玲%王璇%王煜%刘岩%王小晋%张丽%张慧媛%顾德周%王佩荣%陈广全%乐加昌
張捷%許美玲%王璇%王煜%劉巖%王小晉%張麗%張慧媛%顧德週%王珮榮%陳廣全%樂加昌
장첩%허미령%왕선%왕욱%류암%왕소진%장려%장혜원%고덕주%왕패영%진엄전%악가창
食源性病毒%诺如病毒%分子马达生物传感器%快速检测
食源性病毒%諾如病毒%分子馬達生物傳感器%快速檢測
식원성병독%낙여병독%분자마체생물전감기%쾌속검측
food-borne virus%Norovirus%molecular motor biosensor%rapid detection
应用分子马达生物传感技术，建立食品中诺如病毒特异性快速检测方法。以嗜热菌中提取的载色体（chromatophore）为材料，根据诺如病毒ORF1和ORF2连接处的高度保守区设计探针，利用生物素-亲和素系统将诺如病毒探针连接在F0F1-ATPase的ε亚基上构建生物传感器。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，对样品中的RNA进行检测。建立并优化了F0F1-ATPase生物传感技术检测食源性诺如病毒的方法。所建立的方法具有良好的特异性，12个含诺如病毒样品均能检出，3个不含诺如病毒样品均未检出，无假阴性或假阳性情况出现。该方法可在1 h内完成检测，检测灵敏度在RNA水平上达到0.005 ng/mL。所建立的分子马达生物传感技术可以较好的检测食品中的诺如病毒，并为其他病毒的检测提供参考。
應用分子馬達生物傳感技術，建立食品中諾如病毒特異性快速檢測方法。以嗜熱菌中提取的載色體（chromatophore）為材料，根據諾如病毒ORF1和ORF2連接處的高度保守區設計探針，利用生物素-親和素繫統將諾如病毒探針連接在F0F1-ATPase的ε亞基上構建生物傳感器。提取病毒RNA併將其與生物傳感器結閤的同時啟動ATP閤成，比較其熒光彊度的差彆，對樣品中的RNA進行檢測。建立併優化瞭F0F1-ATPase生物傳感技術檢測食源性諾如病毒的方法。所建立的方法具有良好的特異性，12箇含諾如病毒樣品均能檢齣，3箇不含諾如病毒樣品均未檢齣，無假陰性或假暘性情況齣現。該方法可在1 h內完成檢測，檢測靈敏度在RNA水平上達到0.005 ng/mL。所建立的分子馬達生物傳感技術可以較好的檢測食品中的諾如病毒，併為其他病毒的檢測提供參攷。
응용분자마체생물전감기술，건립식품중낙여병독특이성쾌속검측방법。이기열균중제취적재색체（chromatophore）위재료，근거낙여병독ORF1화ORF2련접처적고도보수구설계탐침，이용생물소-친화소계통장낙여병독탐침련접재F0F1-ATPase적ε아기상구건생물전감기。제취병독RNA병장기여생물전감기결합적동시계동ATP합성，비교기형광강도적차별，대양품중적RNA진행검측。건립병우화료F0F1-ATPase생물전감기술검측식원성낙여병독적방법。소건립적방법구유량호적특이성，12개함낙여병독양품균능검출，3개불함낙여병독양품균미검출，무가음성혹가양성정황출현。해방법가재1 h내완성검측，검측령민도재RNA수평상체도0.005 ng/mL。소건립적분자마체생물전감기술가이교호적검측식품중적낙여병독，병위기타병독적검측제공삼고。
This study is to establish rapid detection method for Norovirus in food using molecular motor biosensor technologies. Chromatophores are inverted plasma membrane vesicles containing F0F1-ATPase, and the chromatophores were used as a model system to detect Norovirus. A specific probe was encompassed on the conservative region between ORF1 and ORF2. A biosensors were constructed by connecting specific molecular probes with F0F1-ATPase'sεsubunit throughavidin-biotin system. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP were synthesized. By comparing their fluorescence intensity, we can detect Norovirus RNA in samples. Through optimizing the reaction condition, a simple and rapid method for detection of Noroviris was developed by F0F1-ATPase biosensor. The results of the study indicated that the method had perfect specificity, and analysis of Hepatitis A visus and Rotavirus demonstrated that this system was specific. The test could be done within 1 h with the sensitivity low to 0.005 ng/mL with genomic RNA. Fifteen samples artificially contaminated and fifty samples were detected, and no false positive or false negative results. Result indicated that this reliable, sensitive, quick and easy-handling method could be used for the detection of food-borne Norovirus virus, and could also be a valuable reference to other food-borne viruses.
