摘要
文章结合叠氮溴化丙啶(PMA)处理、羟基萘酚蓝(HNB)和环介导恒温扩增(LAMP)技术,建立了禽畜肉中沙门氏菌活菌的检验方法。结果表明,经0.1%脱氧胆酸钠(SD)37℃处理20 min,再加入10μg/mL浓度PMA,室温避光孵育10 min,曝光15 min,能抑制10~5 cfu/mL沙门氏菌死菌的DNA扩增。建立的LAMP反应体系添加羟基萘酚蓝的浓度为210μmol/L,纯菌灵敏度可达到10~2 cfu/mL,在人工污染肉制品检出限为10~3 cfu/g。在对20份市售禽畜肉的检测中,建立的活菌可视化LAMP检测方法与国家标准方法GB 4789.4—2016相比,没有出现假阳性的情况。建立的方法为畜禽肉中沙门氏菌的检测提供了新技术,也为其他致病菌的检测提供了新思路。
In this paper,a method for the detection of live Salmonella in livestock and poultry meat combining with propidium monoazide(PMA)treatment,hydroxynaphtholblue(HNB)and loop-mediated isothermal amplification technology was established.The results showed that when treated with 0.1%sodium deoxycholate(SD)at 37℃for 20 min,incubated at room temperature for 10 minutes in the dark in concentration of 10μg/mL PMA,and then exposed for 15 minutes,DNA amplification of 10~5 cfu/mL dead Salmonella cells could be inhibited.A final HNB concentration of 210μmol/L was added in the LAPM system.The pure bacterial solution sensitivity of the method was 10~2 cfu/mL,while the detection limitin artificially contaminated meat was 10~3 cfu/mL.In the inspection of 20 marketed livestock and poultry meat,there was no false positive result in the method established compared with national standard method(GB 4789.4—2016).The established method provided a new technology for the detection of Salmonella in livestock and poultry meat,and also provided a new idea for the detection of other pathogenic bacteria.
作者
张珣
戴小芳
曾慧君
董华夏
付诗慧
郦娟
ZHANG Xun;DAI Xiaofang;ZENG Huijun;DONG Huaxia;FU Shihui;LI Juan(Hubei Academy of Agricultural Sciences Institute of Agricultural Quality Standards and Testing Technology,Wuhan 430064,China;Wuhan Institute for Food and Cosmetic Control,Wuhan 430012,China)
出处
《食品科技》
CAS
北大核心
2020年第5期350-355,共6页
Food Science and Technology
基金
湖北省自然科学基金计划项目(2017CFB366)
江苏省海洋药物活性分子筛选重点实验室开放基金项目(HY201702)。
关键词
沙门氏菌
叠氮溴化丙锭
羟基萘酚蓝
环介导等温扩增技术
活菌
Salmonella
propidiummonoazide
hydroxynaphthol blue
loop-mediated isothermal amplification
live cell
