4种食源性致病菌的多重PCR快速检测方法研究  

Study on Multiplex PCR of Rapid Detection Method for Four Foodborne Pathogens

作　　者：李聪[1] 刘健慧 李志辉[1] 张敏[1] 高浩 檀建新[1] LI Cong;LIU Jian-hui;LI Zhi-hui;ZHANG Min;GAO Hao;TAN Jian-xin(College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,Hebei,China)

机构地区：[1]河北农业大学食品科技学院

出　　处：《食品研究与开发》2019年第21期164-169,共6页Food Research and Development

基　　金：河北省重点研发计划项目(16275505D)

摘　　要：为快速检测蜡样芽孢杆菌、金黄色葡萄球菌、志贺氏菌和沙门氏菌4种食源性致病菌,建立4重聚合酶链式反应(polymerase chain reaction,PCR)的检测方法。针对菌株的特异性基因设计引物,选择退火温度相近,扩增条带区间不同的引物为多重聚合酶链式反应引物组,并对多重PCR的引物浓度、退火温度进行优化,确定扩增条件。结果表明,在54℃的退火温度下,4种菌可扩增长度为246、166、65、107 bp的清晰片段,其检测灵敏度为蜡样芽孢杆菌2×10^1 CFU/mL,金黄色葡萄球菌可达1.3×10^2 CFU/mL,志贺氏菌可达1.3×10^2 CFU/mL,沙门氏菌可达1.4×10^2 CFU/mL。A multiplex polymerase chain reaction(PCR)method for rapid detection of four foodborne pathogens,Bacillus cereus,Staphylococcus aureus,Shigella and Salmonella was established.Four pair of primers were designed for detection of the specific genes of the strains.The primers with similar annealing temperature and different amplification bands were selected to be PCR primers.The primer concentration and annealing temperature were optimized to determine the amplification conditions.Result showed when the annealing temperature of 54℃,the four strains could expand 246,166,65,107 bp of clear fragments.The detection sensitivity was 2×10^1 CFU/mL for Bacillus cereus,1.3×10^2 CFU/mL for Staphylococcus aureus,1.3×10^2 CFU/mL for Shigella and 1.4×10^2 CFU/mL for Salmonella.

关 键 词：多重聚合酶链式反应 蜡样芽孢杆菌 金黄色葡萄球菌 志贺氏菌 沙门氏菌 快速检测 

分 类 号：S85[农业科学—兽医学]