河北工业科技2024，Vol.41Issue(2)：133-140,8.DOI:10.7535/hbgykj.2024yx02007
4种常见食源性病原菌多重PCR检测技术研究
Study on multiple PCR detection techniques for four common foodborne pathogens
摘要
Abstract
The method of efficient multiplex polymerase chain reaction(MPCR)was constructed based on the amplification of specific gene sequences of four common food borne pathogens in order to solve the problems of those traditional detection methods involving cumbersome operations and time consuming.The MPCR method was established using the designed primers according to the specific sequences of iap gene in Listeria monocytogenes,gyrB gene in Bacillus cereus,stxⅠ gene in Shiga toxin-producing Escherichia coli,invA gene in Salmonella enteritidis,respectively.The specificity,the sensitivity and the feasibility of MPCR were analyzed,and it was compared with the national standard culture method.The results show that the specific bands of four target genes amplified by MPCR were 371,221,432 and 171 bp,respectively.Under the annealing temperature of 58℃,the MPCR presented strong specificity without non-specific amplification,and the lowest detection limit(LOD)of four pathogenic bacteria reached up to 102 CFU/mL.The two detecting results of the MPCR and the standard culture method were identical,and the detection period of MPCR was reduced to 8～9 h from that of traditional methods for 5～7 d.The constructed MPCR,as a rapid and efficient detecting method for food products in practice,provided guarantees for food safety.关键词
食品检验学/多重PCR(MPCR)/单增李斯特菌/蜡样芽孢杆菌/产志贺毒素大肠埃希氏菌/肠炎沙门氏菌Key words
food inspection/multiplex PCR(MPCR)/Listeria monocytogenes/Bacillus cereus/Shiga toxin-producing Escherichia coli/Salmonella enteritidis分类
轻工纺织引用本文复制引用
王珊,王雁伟,高辉明,艾鹏飞..4种常见食源性病原菌多重PCR检测技术研究[J].河北工业科技,2024,41(2):133-140,8.基金项目
河北省科技计划"奶业振兴重大技术创新专项"(19227137D) （19227137D）
河北省大中学生科技创新专项(22E50074D) （22E50074D）
