Multiplex PCR for simultaneous identification of E. coli O157:H7, Salmonella spp. and L. monocytogenes in food.
The rapid detection of pathogens in food is becoming increasingly critical for ensuring the safety of consumers, since the majority of food-borne illnesses and deaths are caused by pathogenic bacteria. Hence, rapid, sensitive, inexpensive and convenient approaches to detect food-borne pathogenic bacteria is essential in controlling food safety. In this study, a multiplex PCR assay for the rapid and simultaneous detection of Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes was established. The invA, stx and hlyA genes specifically amplified DNA fragments of 284, 404 and 510 bp from Salmonella spp., L. monocytogenes and E. coli O157:H7, respectively. The 16S rRNA gene was targeted as an internal control gene in the presence of bacterial DNA. The specificity and sensitivity of the multiplex PCR were performed by testing different strains. The multiplex PCR assay was able to specifically simultaneously detect ten colony-forming unit/mL of each pathogen in artificially inoculated samples after enrichment for 12 h. The whole process took less than 24 h to complete, indicating that the assay is suitable for reliable and rapid identification of these three food-borne pathogens, which could be suitable in microbial epidemiology investigation.
Department of Pharmacy, Ho Chi Minh City University of Technology (HUTECH), 475A Dien Bien Phu Street, Ward 25, Binh Thanh District, Ho Chi Minh City, Vietnam.|Department of BionanoTechnology, Gachon Medical Research Institute, Gachon University, Sungnam, Korea.
Thuy Trang
TT
Nguyen
Department of Faculty of Food Technology, Ho Chi Minh City University of Food Industry (HUFI), 140 Le Trong Tan, Tan Phu district, Ho Chi Minh City, Vietnam. vovangiau911@gmail.com.|Department of BionanoTechnology, Gachon Medical Research Institute, Gachon University, Sungnam, Korea. vovangiau911@gmail.com.
Vo
V
Van Giau
Vietnam Sports Hospital, Ministry of Culture, Sports and Tourism, Do Xuan Hop Road, My Dinh I Ward, Nam Tu Liem District, Hanoi, Vietnam. votuongkha@yahoo.com.
Tuong Kha
TK
Vo
Detection; Food-borne pathogen; Multiplex PCR; Simultaneous