食品科学 ›› 2011, Vol. 32 ›› Issue (15): 308-311.doi: 10.7506/spkx1002-6630-201115067

• 专题论述 • 上一篇    下一篇

细菌活的非可培养状态的分子生物学检测技术研究进展

李正义,贾俊涛*,祝素珍,姜英辉,房保海,雷质文   

  1. 山东出入境检验检疫局
  • 出版日期:2011-08-15 发布日期:2011-07-26
  • 基金资助:
    国家质检总局科研项目(2010IK180)

Recent Advances in Studies on Molecular Biology Methods for Detecting Bacteria in Viable but Nonculturable State

LI Zheng-yi,JIA Jun-tao*,ZHU Su-zhen,JIANG Ying-hui,FANG Bao-hai,LEI Zhi-wen   

  1. (Shandong Entry-Exit Inspection and Quarantine Bureau, Qingdao 266002, China)
  • Online:2011-08-15 Published:2011-07-26

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。

关键词: 细菌, 活的非可培养状态, 分子生物学, 检测技术

Abstract: 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.

Key words: bacteria, viable but nonculturable state, molecular biology, detection technique

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