应用PMA-qPCR方法快速准确检测发酵乳制品中副干酪乳杆菌活菌的研究  

Development of PMA-qPCR assay for rapid and accurate detection of viable Lactobacillus paracasei in fermented dairy products

作　　者：王力均[1] 谭强来[1] 朱江 陈奇[1] 李萍[1] 叶若松[1] 徐锋[1] 魏华[1] 许恒毅[1] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]江西省食品工业研究所,江西南昌330029

出　　处：《中国微生态学杂志》2013年第1期1-4,共4页Chinese Journal of Microecology

基　　金：国家自然基金项目(31170091;31000048;30900038);食品科学与技术国家重点实验室项目(SKLF-TS-200916;SKLF-MB-201002)

摘　　要：目的建立快速、准确检测发酵乳制品中副干酪乳杆菌活菌的方法。方法发酵乳制品中副干酪乳杆菌先经PMA处理后,采用沸水浴的方法提取副干酪乳杆菌基因组,然后通过qPCR方法检测发酵乳制品中的活菌。结果副干酪乳杆菌经90℃处理6 min,即为膜损伤菌;PMA能够抑制107CFU/mL死菌DNA的扩增,而不影响活菌DNA的扩增;PMA-qPCR能够准确检测到样品中活菌。结论建立了一种快速、准确的方法检测发酵乳制品中的副干酪乳杆菌活菌。Objective To develop a rapid and accurate method for detection of viable Lactobacillus paracasei in fermented dairy products.Methods L.paracasei in fermented dairy products was treated with PMA（propidium monoazide） prior to the extraction of DNA by boiling.Then viable L.paracasei in fermented dairy products was quantified by qPCR.Results After 6 min of incubation at 90 ℃,the membrane of L.paracasei was injured.PMA had inhibited the PCR amplification of DNA from dead cells at the concentration of 107 CFU/mL and did not affect the amplification of DNA from viable cells.The PMA-qPCR assay had accurately detected the viable L.paracasei in fermented dairy products.Conclusion A PMA-qPCR assay has been developed for rapid and accurate detection of viable L.paracasei in fermented dairy products.

关 键 词：副干酪乳杆菌 叠氮溴化丙锭 荧光定量PCR 检测 

分 类 号：Q93-332[生物学—微生物学]