PCR method based on the ogdH gene for the detection of Salmonella spp. from chicken meat samples.
In a previous paper, the ogdH gene that encodes 2-oxoglutarate dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-1 and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method.
Jin
Un-Ho
UH
Department of Biochemistry and Molecular Biology, College of Oriental Medicine, Dongguk University and NRL-Glycobiology, Kyungju 780-714, Korea.
Cho
Sung-Hak
SH
Kim
Min-Gon
MG
Ha
Sang-Do
SD
Kim
Keun-Sung
KS
Lee
Kyu-Ho
KH
Kim
Kwang-Yup
KY
Chung
Duck Hwa
DH
Lee
Young-Choon
YC
Kim
Cheorl-Ho
CH
D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000382:microbiology*; D015169:Colony Count, Microbial; D017931:DNA Primers; D007655:Ketoglutarate Dehydrogenase Complex / Q000235:genetics* / Q000378:metabolism; D008460:Meat / Q000382:microbiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012475:Salmonella / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D007655:Ketoglutarate Dehydrogenase Complex