PMA-ddPCR技术定量检测VBNC态沙门氏菌  

A Quantitative Assay for Measuring VBNC Salmonella by Droplet Digital PCR Combined with Propidium Monoazide

作　　者：冯秋实 程逸宇 吴海晶 周海波 沈威 唐思颉 刘新梅 FENG Qiushi;CHENG Yiyu;WU Hajing;ZHOU Haibo;SHEN Wei;TANG Sijie;LIU Xinmei(Nanjing Institute for Food and Drug Control,Nanjing 211198,China)

机构地区：[1]南京市食品药品监督检验院,江苏南京211198

出　　处：《食品科技》2024年第7期336-342,共7页Food Science and Technology

基　　金：国家市场监督管理总局科技计划项目(2022MK158);江苏省市场监督管理局科技计划项目(KJ2024017)。

摘　　要：为实现活的非可培养(Viable but nonculturable state, VBNC)态沙门氏菌的定量检测,文章根据沙门氏菌的特异性毒力基因设计引物和探针,优化微滴数字聚合酶链反应(Droplet digital polymerase chain reaction, ddPCR)反应条件及叠氮丙啶(Propidium monoazide, PMA)处理方式,构建PMA-ddPCR快速定量检测方法。结果表明,拷贝数与菌液浓度在10^(3)~10^(7) CFU/mL范围内呈现良好的线性关系,PMA-ddPCR方法检出限为10^(2) CFU/mL,定量限为10^(3) CFU/mL。所建立的PMA-ddPCR快速定量检测冷冻肉制品中VBNC态沙门氏菌方法,弥补了现有标准检测方法的不足。In order to quantitatively detect viable but nonculturable state(VBNC)Salmonella,primers and probes were designed based on the specific virulence gene of Salmonella,and droplet digital polymerase chain reaction(ddPCR)reaction conditions and propidium monoazide(PMA)treatment were optimized.A rapid quantitative detection method of PMA-ddPCR was established.The results showed a good linear relationship between copy number and bacterial solution concentration in the range of 10^(3)–10^(7) CFU/mL.The detection limit and quantitation limit of PMA-ddPCR were 10^(2) CFU/mL and 10^(3) CFU/mL respectively.The established PMA-ddPCR method for rapid and quantitative detection of VBNC Salmonella in frozen meat products addresses the shortcomings of existing standard detection methods.

关 键 词：叠氮丙啶 微滴式数字PCR 沙门氏菌 活的非可培养态 定量检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]