Apirak VisetsripongKobchai PattaragulwanitJiraporn ThaniyavarnRyosuke MatsuuraAkio KurodaOrasa SutheinkulChulalongkorn UniversityKyoto Biseibutsu InstituteKyoto City Institute of Health and Environmental SciencesMahidol University2018-08-242018-08-242007-01-01Southeast Asian Journal of Tropical Medicine and Public Health. Vol.38, No.1 (2007), 82-90012515622-s2.0-33947584403https://repository.li.mahidol.ac.th/handle/20.500.14594/25064A rapid method for detection of Escherichia coli O157: H7 using multiplex PCR was developed. Two oligonucleotide primer pairs were used for simultaneously detection of vt encoding verotoxin genes for virulence factor and rfb O157 encoding the O-antigen specific for E. coli O157: H7. Multiplex PCR generated two products of 215 bp and 420 bp for vt and rfbO157, respectively. Multiplex PCR detected reference strain O157: H7 (NF-7777) with a sensitivity of 105 CFU per ml with no amplification of other 15 pathogenic bacteria. After incubation of 102 CFU/25 gram raw meat in tryptic soy broth at 37°C for 8 hours, multiplex PCR conducted with the addition of 100 mg bovine serum albumin produced the two specific PCR products for E. coli O157: H7. This modified multiplex PCR is a rapid, sensitive, and specific technique for detecting and differentiating E. coli O157: H7 and has the potential to be used as an alternative to conventional methods for the screening of O157: H7 strains isolated from raw meat.Mahidol UniversityMedicineArticleSCOPUS