摘要
许多细菌在不良环境下能进入活的非可培养状态(viable but nonculturable state,VBNC)。细菌培养技术常常造成VBNC状态的细菌漏检,应用分子生物学检测技术可以提高VBNC细胞的阳性检出率。针对VBNC细胞的分子生物学检测技术,基于细菌特异性DNA分子、mRNA分子是目前常用的检测方法,而利用报告基因(如绿色荧光蛋白基因)检测VBNC细胞是一种有效的方法。最近利用叠氮溴乙锭(ethidium monoazide bromide,EMA)或者叠氮溴化丙锭(propidium monoazide,PMA)联合PCR技术选择性抑制细菌死细胞扩增,该方法结合了EMA(PMA)选择渗透性和PCR特异性,作为一种区分细胞死活的方法,可以有效检测细菌VBNC细胞。
A number of bacteria have been reported to have the capability to enter a viable but nonculturable（VBNC） state as a response to environmental stress.Cell culture techniques often fail to detect bacterial cells in the VBNC state;however,molecular biology methods can improve the detection efficiency for VBNC cells.Among molecular biology methods for detecting bacterial cells in the VBNC state,the detection of specific DNA or mRNA is a common method and a gfp gene encoding green fluorescent protein has been used as an effective marker to examine VBNC state.Recently,a combinatorial method,designated as ethidium monoazide（EMA）-PCR method,has also been established to detect live/dead cells due to the selective penetration of EMA/PMA into dead cells in the presence of live cells.This detection method combined with the selectivity of EMA/PMA and the specificity of PCR is sufficient to detect viable cells and VBNC cells.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2011年第15期308-311,共4页
Food Science
基金
国家质检总局科研项目(2010IK180)
关键词
细菌
活的非可培养状态
分子生物学
检测技术
bacteria
viable but nonculturable state
molecular biology
detection technique
