Simultaneous detection of viable Salmonella spp., Escherichia coli, and Staphylococcus aureus in bird's nest, donkey-hide gelatin, and wolfberry using PMA with multiplex real-time quantitative PCR

Taobo Liang; Hui Long; Zhongxu Zhan; Yingfei Zhu; Peilin Kuang; Ni Mo; Yuping Wang; Shenghui Cui; Xin Wu

doi:10.1002/fsn3.2916

Simultaneous detection of viable Salmonella spp., Escherichia coli, and Staphylococcus aureus in bird's nest, donkey-hide gelatin, and wolfberry using PMA with multiplex real-time quantitative PCR

Food Sci Nutr. 2022 May 17;10(9):3165-3174. doi: 10.1002/fsn3.2916. eCollection 2022 Sep.

Authors

Taobo Liang¹, Hui Long², Zhongxu Zhan¹, Yingfei Zhu¹, Peilin Kuang¹, Ni Mo¹, Yuping Wang³, Shenghui Cui⁴, Xin Wu¹

Affiliations

¹ Jiangxi Institute for Food Control Nanchang China.
² Nanchang Center for Disease Control and Prevention Nanchang China.
³ Chengdu Institute of Food and Drug Control Chengdu China.
⁴ National Institutes for Food and Drug Control Beijing China.

Abstract

Salmonella spp., Escherichia coli, and Staphylococcus aureus are common microbial contaminants within the homology of medicine and food that can cause serious food poisoning. This study describes a highly efficient, sensitive, specific, and simple multiplex real-time quantitative PCR (mRT-qPCR) method for the simultaneous detection of viable Salmonella spp., E. coli, and S. aureus. Primers and probes were designed for the amplification of the target genes invA, uidA, and nuc. Dead bacterial genetic material was excluded by propidium monoazide (PMA) treatment, facilitating the detection of only viable bacteria. This method was capable of detecting Salmonella spp., E. coli, and S. aureus at 10², 10², and 10¹ CFU/ml, respectively, in pure culture. PMA combined with mRT-qPCR can reliably distinguish between dead and viable bacteria with recovery rates from 95.7% to 105.6%. This PMA-mRT-qPCR technique is a highly sensitive and specific method for the simultaneous detection of three pathogens within the homology of medicine and food.

Keywords: homology of medicine and food; mRT‐qPCR; pathogens; viable bacteria detection.