食源性致病菌多重PCR检测技术建立及其应用

胡金强; 丁慧敏; 詹丽娟; 赵卫东; 侯莹莹; 孙新城; 高辉; 耿尧

doi:10.12187/2022.01.002

Volume 37 Issue 1

March 2022

Article Contents

Abstract

References

HU Jinqiang, DING Huimin, ZHAN Lijuan, et al. Development and application of multiplex PCR detection techniques for foodborne pathogenic bacteria[J]. Journal of Light Industry, 2022, 37(1): 12-19. doi: 10.12187/2022.01.002

Citation: HU Jinqiang, DING Huimin, ZHAN Lijuan, et al. Development and application of multiplex PCR detection techniques for foodborne pathogenic bacteria[J]. Journal of Light Industry, 2022, 37(1): 12-19. doi: 10.12187/2022.01.002 shu

Development and application of multiplex PCR detection techniques for foodborne pathogenic bacteria

HU Jinqiang ^1,2,3 ,
DING Huimin ¹ ,
ZHAN Lijuan ⁴ ,
ZHAO Weidong ¹ ,
HOU Yingying ¹ ,
SUN Xincheng ¹ ,
GAO Hui ¹ ,
GENG Yao ¹

1. College of Food and Bioengineering, Zhengzhou University of Light Industry, Zhengzhou 450001, China;
2. International Joint Laboratory of Food Safety of He'nan Province, Zhengzhou 450001, China;
3. Collaborative Innovation Center of Food Production and Safety of He'nan Province, Zhengzhou 450001, China;
4. College of Food Science and Technology, He'nan Agricultural University, Zhengzhou 450002, China

Received Date: 2021-05-07

Abstract

Multiple PCR detection techniques were established for 6 foodborne pathogenic bacteria specific genes (the nuc gene of Staphylococcus aureus, tdh gene of Vibrio parahaemolyticus, hly gene of Listeria monocytogenes, ompA gene of Cronobacter sakazakii, invA gene of Salmonella typhimurium, and rfbE gene of Escherichia coli O157:H7). The sensitivity and specificity of multiplex PCR were analyzed, and its application feasibility in artificially-contaminated milk was also evaluated. The results showed that the specific PCR products of 345 bp, 307 bp, 269 bp, 237 bp, 188 bp and 142 bp were amplified as expected in multiplex PCR without nonspecific amplification and cross reactivity with non-target pathogenic bacteria. Multiplex PCR developed in this study could detect as little as 10 pg of genomic DNA of pure bacterial culture and 10³ CFU/mL of milk artificially-contaminated with pathogenic bacteria, which could specifically, sensitively, rapidly and efficiently detect six foodborne pathogenic bacteria simultaneously.
- foodborne pathogenic bacteria,
- multiplex PCR,
- detection techniques
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