微生态水样中活的非可培养态大肠杆菌O157：H7的检测  

Detection of the viable but nonculturable Escherichia coil O157：H7 in aquatic microcosm

作　　者：罗中捷[1,2] 周卓晟[1,3] 郝春慧[1] 郭寅生[1] 刘红艳[1] 郑华英[4] 黄正[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院环境与健康教育部重点实验室,武汉430030 [2]广州市卫生监督所 [3]深圳市龙岗区龙岗卫生监督分所 [4]武汉市疾病预防控制中心

出　　处：《中华预防医学杂志》2012年第2期129-132,共4页Chinese Journal of Preventive Medicine

摘　　要：目的监测大肠杆菌O157：H7在微生态汉江水中活的非可培养态；探讨荧光染色、异养平板计数和ELISA检测活的非可培养态大肠杆菌O157：H7的可行性。方法将大肠杆菌O157：H7投加到模拟微生态汉江水样中，通过改变温度和营养条件，分别采用吖啶橙染色、活菌直接计数-5-氰基-2，3-二-（P-苄基-四唑氯化物）染色、活菌直接计数·萘啶酮酸染色、异养平板计数和ELISA法实时监测大肠杆菌O157：H7的存活情况。结果在4℃、寡营养条件下，大肠杆菌O157：H7在汉江水中存活58d后进入活的非可培养态，此时采用活菌直接计数-5-氰基-2，3-二-（P-苄基-四唑氯化物）染色法测得活菌数为1．2×10^5CFU／ml，活菌直接计数-萘啶酮酸染色法测得活菌数为9．0×10^4CFU／ml，异养乎板计数法测得可培养菌数为0，ELISA法测得微生态汉江水中大肠杆菌O157：H7总菌数在58d内基本稳定于10^6CFU／ml的初始菌数水平。结论在4℃、寡营养条件下，大肠杆菌O157：H7在微生态汉江水中可进入活的非可培养态；ELISA结合荧光染色、异养平板计数，能定量检出活的非可培养态大肠杆菌O157：H7。Objectives This study is to monitor the survival of E. coli O157：H7 in the aquatic microcosm from Han River and explore the feasibility of fluorescence staining, heterotrophic plate count and ELISA in detection of viable but nonculturable E. coli O157： H7. Methods E. coli O157：H7 were added into aquatic microcosm from Han River and cultured with low temperature and olig-nutrition. Then the survival of E. coli O157：H7 were real-time monitored by acridine orange direct count （AODC） , direct viable count （DVC）-CTC staining, DVC-nalidixic acid （NA） staining, heterotrophic plate count （HPC） and ELISA. Results E. coli O157： H7 can be converted to a viable hut nonculturable state in the aquatic microcosm from Han River 58 days after cultured at 4 ℃ with oligo-nutrition. The amount of viable E. coli O157：H7 was measured as 1.2 × 10^5 CFU/ml by DVC-CTC and 9. 0 × 10^4 CFU/ml by DVC-NA, whereas the amount of culturable bacterial determined by HPC is 0. The amounts of bacteria determined by ELISA are basically stable within 58 days around 10^6 CFU/ml. Conclusion E. coli O157：H7 can be converted into a viable but nonculturable state in Han River water at 4 ℃ with oligo-nutrition, and ELISA combined with fluorescence staining and heterotrophic plate count can be used in quantitative detection of the viable but noncuhurable E. coli O157： H7.

关 键 词：大肠杆菌O157 生态学 酶联免疫吸附测定 活的非可培养态 微生态 

分 类 号：R123.1[医药卫生—环境卫生学]