重组酶介导扩增技术快速检测食品中单核细胞增生李斯特菌  

Rapid detection of Listeria monocytogenes by recombinase aided amplification method

作　　者：崔荣飞 赵义良 田梅 赵永坡[2] 曹楠 王丽娟 郭利川 CUI Rong-Fei;ZHAO Yi-Liang;TIAN Mei;ZHAO Yong-Po;CAO Nan;WANG Li-Juan;GUO Li-Chuan(Shijiazhuang Animal Products and Veterinary Medicine Feed Quality Testing Center,Shijiazhuang 050041,China;Shijiazhuang Academy of Agricultural and Forestry Sciences,Shijiazhuang 050000,China;Institute of Animal Health Supervision,Shijiazhuang 050000,China;Jiangsu Qitian Gene Biotechnology Co.,Ltd,Wuxi 214000,China)

机构地区：[1]石家庄市畜产品和兽药饲料质量检测中心,石家庄050041 [2]石家庄市农林科学研究院,石家庄050000 [3]河北省动物卫生监督所,石家庄050000 [4]江苏奇天基因生物科技有限公司,无锡214000

出　　处：《食品安全质量检测学报》2021年第5期1773-1777,共5页Journal of Food Safety and Quality

基　　金：河北省重点研发计划项目(19227503D)。

摘　　要：目的建立重组酶介导扩增技术(recombinase aided amplification,RAA)快速检测单核细胞增生李斯特菌的方法。方法以单核细胞增生李斯特菌的hlyA基因作为靶序列,设计引物和探针,通过构建含目的基因片段的重组质粒,评价RAA方法的灵敏度,通过检测沙门氏菌核酸、金黄色葡萄球菌核酸、副溶血性弧菌核酸、溶血性链球菌核酸、志贺氏菌核酸基因组DNA,评价其特异性。结果建立的RAA方法可在39℃、20 min内,检测重组质粒中hlyA基因片段,最低可检出的质粒拷贝数为10拷贝/反应,且与沙门氏菌、金黄色葡萄球菌、副溶血性弧菌、溶血性链球菌、志贺氏菌基因组DNA无交叉反应。结论建立的RAA方法具有灵敏度高和特异性强的特点,可用于单核细胞增生李斯特菌的快速检测。Objective To establish a recombinase aided amplification(RAA)method for detection of Listeria monocytogenes.Methods The primers and probe were designed according the specific hlyA gene sequence of Listeria monocytogenes.The sensitivity of RAA method was evaluated using a recombinant plasmid containing hlyA gene fragment.The specificity was performed by amplification of genomic DNA of Salmonella,Staphylococcus aureus,Vibrio Parahemolyticus,Streptococcus hemolyticus and Shigella.Results RAA method can be used to detect hlyA gene fragment in the recombinant plasmid at a copy number as low as 10 copies per reaction within 20 min at 39℃with no cross-reaction with Salmonella,Staphylococcus aureus,Vibrio Parahemolyticus,Streptococcus hemolyticus and Shigella.Conclusion RAA method has the characteristics of high sensitivity and specificity,and can be used for rapid detection of Listeria monocytogenes.

关 键 词：单核细胞增生李斯特菌 重组酶介导扩增 分子检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]