多管发酵-菌落PCR法定量检测水中可培养沙门氏菌  

Quantitative Detection of Culturable Salmonella in Water by Multiple Tube Fermentation Combined with Colony PCR

作　　者：张崇淼[1] 王晓昌[1] 周进宏[1] 孙婷婷[1] 魏梦楠[1] 曾颂[1] 

机构地区：[1]西安建筑科技大学环境与市政工程学院,西北水资源与环境生态教育部重点实验室,西安710055

出　　处：《应用与环境生物学报》2012年第6期1004-1008,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(No.50908185);中国博士后基金项目(No.20100470086);陕西省教育厅自然科学研究项目(No.11JK0765)~~

摘　　要：采用膜吸附-洗脱法浓缩水样,通过多管发酵法筛选出以可培养沙门氏菌为主的活体细菌.根据沙门氏菌invA基因的保守性,利用沙门氏菌属通用引物进行菌落PCR鉴定,进而确定沙门氏菌阳性平板数量.运用最大可能数法分析数据,计算出水样中可培养沙门氏菌的含量.该方法对沙门氏菌具有良好的特异性和涵盖性.通过对人工污染水样和城市污水厂二级处理出水的检测,证明了多管发酵-菌落PCR方法的检测结果较为准确,可操作性强,能够更真实地反映样品中沙门氏菌的活性.To establish a quantitative detection method of culturable Salmonella in water sample, the multiple tube fermentation （MTF） was used to preliminarily select the viable bacteria in which culturable Salmonella are predominant after water concentration by membrane adsorption - elution method. According to the conserved region of invA gene, Salmonella positive strain was identified by colony - PCR with universal primer pairs. The concentration of Salmonella in water sample was calculated by most probably number （MPN） method. This method has good specificity to all Salmonella species, and no cross-reactivity was observed using the genome of other bacteria as PCR template. The validation data of the artificial polluted water and secondary effluent samples indicated that MTF-colony PCR method was accurate, labor-saving and could reflect the activity of Salmonella in water samples.

关 键 词：可培养沙门氏菌 多管发酵-菌落PCR法 膜吸附-洗脱法 定量检测 最大可能数法 

分 类 号：X832[环境科学与工程—环境工程]