| 万志刚1 汤慕瑾1 吕敬章1 罗志军2 洪小柳1 马淑棉1.多种食源性致病菌检测的多重PCR 方法的研究[J].,2012,12(11):2177-2181 |
| 多种食源性致病菌检测的多重PCR 方法的研究 |
| Study of A Multiplex PCR Method for the Detection of Foodborne Pathogen |
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| DOI： |
| 中文关键词: 多重PCR 食源性致病菌 |
| 英文关键词: Multiplex PCR Food borne pathogen |
| 基金项目:深圳出入境检验检疫局科技项目（SZ2007004） |
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| 中文摘要: |
| 目的：利用多重PCR 技术，建立可以同时检测多种食源性致病菌的多重PCR 方法。方法：分别选择沙门氏菌invA 基因，志
贺氏菌的ipaH 基因，单核细胞增生李斯特氏菌的hlyA 基因，大肠杆菌O157：H7 的eaeA 基因，副溶血弧菌的toxR 基因，设计多
重PCR 引物，建立多重PCR 检测体系，并对该体系进行特异性和灵敏度实验。结果：通过对19 株菌株进行实验，所有的目标菌株
均为阳性，而其余菌株为阴性。对多重PCR 体系的灵敏度进行考察，沙门菌的灵敏度为5000 CFU/mL；志贺氏菌的灵敏度为5500
CFU/mL；单核细胞增生李斯特氏菌的灵敏度为5200 CFU/mL；O157：H7 的灵敏度为5000CFU/mL；副溶血弧菌的灵敏度为6300
CFU/mL。结论：建立的多重PCR 体系能实现多种致病菌同时检测。 |
| 英文摘要: |
| Objective: To develop a multiplex PCR method to detect five food borne pathogenic microorganisms simultaneously.
Methods: Primers specific for invA gene of Salmonella spp., ipaH gene of Shigella spp., hlyA gene of Listeria monocytogenes, eaeA gene
of Escherichia coli O157:H7 and toxR gene of Vibrio parahaemolyticus were designed, and the specificity and sensitivity of the developed
method was further verified. Results: A collection of 19 strains was examined, all target strains were detected. In contrast, none of
the non-target strains yielded the specific amplification product. The sensitivity of the multiplex PCR system was 5000 CFU/mL for
Salmonella cultures, 5500 CFU/mL for Shigella cultures, 5200 CFU/mL for Listeria monocytogenes cultures, 5000 CFU/mL for Escherichia
coli O157:H7 cultures and 6300 CFU/mL for Vibrio parahaemolyticus cultures. Conclusions: The multiplex PCR method in
present study can be applied in practice. |
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