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投稿时间：2018-12-11
投稿时间：2018-12-11
中文摘要: 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2×101 CFU/mL，金黄色葡萄球菌可达 1.3×102 CFU/mL，志贺氏菌可达 1.3×102 CFU/mL，沙门氏菌可达 1.4×102 CFU/mL。
Abstract: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 ×101 CFU/mL for Bacillus cereus，1.3 ×102 CFU/mL for Staphylococcus aureus，1.3 ×102 CFU/mL for Shigella and 1.4×102 CFU/mL for Salmonella.
keywords: multiplex polymerase chain reaction Bacillus cereus Staphylococcus aureus Shigella Salmonella rapid detection
文章编号：201921028 中图分类号： 文献标志码：
基金项目:河北省重点研发计划项目（16275505D）。
| Author Name | Affiliation |
| 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 |
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