DNA染料对死活细胞区别检测的应用研究进展.pdfVIP

中国科技论文在线 DNA 染料对死活细胞区别检测的应用研究 进展# 刘艳艳，宋杰，李岩松，任洪林，柳增善，卢士英** 5 （吉林大学人兽共患病研究所，人兽共患病研究教育部重点实验室，长春 130062 ） 摘要：细菌的可培养性是认定细菌是否存活的传统判定标准，现今活细胞的检测技术已经趋 向于检测细胞的反应性、生理代谢活动、能量状态等。光敏染料叠氮溴化丙锭（PMA ）和 叠氮溴化乙锭（EMA ）是两种能渗透进入膜破损的细胞内与DNA 结合、使DNA 发生不可 逆的修饰作用、抑制DNA 扩增的染料。样品首先用染料进行预处理，随后结合荧光定量PCR 10 （QPCR ）技术，不仅可以对样品中的活细菌进行检测，也可用于检测样品中的真菌、病毒 等。尽管该技术已广泛应用，但在环境样品检测时还存在许多局限。本文总结了光敏染料的 发展状况，并提出了其在应用时应注意的一些关键问题。 关键词：活细胞检测；PMA；EMA；荧光定量PCR 中图分类号：X172 15 Progess of dyes combinating with DNA in detection of live cells LIU Yanyan, SONG Jie, LI Yansong, REN Honglin, LIU Zengshan, LU Shiying (Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, Jilin 20 University, ChangChun 130062) Abstract: Bacteria were traditionally considered viable when they could be cultured, whereas today’s viability concept tends to be based on the presence of some form of responsiveness, metabolic activity, the energy status. Microbiological samples pretreatment with nucleic acid dye PMA and EMA that selectively enters cells with compromised cell membranes, whereas an intact 25 membrane presents a barrier for these molecule. Once inside the dead cells, the dye intercalates into the cell’s DNA which results in irreversible DNA modification and subsequent inhibition of its amplification. The principle is not limited to bacteria, but has also successfully been applied to fungi, protozoa and viruses, Despite the success of the method, some