Identification of Salmonella enterica serovar Typhimurium using specific PCR primers obtained by comparative genomics in Salmonella serovars.
Salmonella enterica serovar Typhimurium is a major foodborne pathogen throughout the world. Until now, the specific target genes for the detection and identification of serovar Typhimurium have not been developed. To determine the specific probes for serovar Typhimurium, the genes of serovar Typhimurium LT2 that were expected to be unique were selected with the BLAST (Basic Local Alignment Search Tool) program within GenBank. The selected genes were compared with 11 genomic sequences of various Salmonella serovars by BLAST. Of these selected genes, 10 were expected to be specific to serovar Typhimurium and were not related to virulence factor genes of Salmonella pathogenicity island or to genes of the O and H antigens of Salmonella. Primers for the 10 selected genes were constructed, and PCRs were evaluated with various genomic DNAs of Salmonella and non-Salmonella strains for the specific identification of Salmonella serovar Typhimurium. Among all the primer sets for the 10 genes, STM4497 showed the highest degree of specificity to serovar Typhimurium. In this study, a specific primer set for Salmonella serovar Typhimurium was developed on the basis of the comparison of genomic sequences between Salmonella serovars and was validated with PCR. This method of comparative genomics to select target genes or sequences can be applied to the specific detection of microorganisms.
Kim
H J
HJ
Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University, Suwon, 449-701, Korea.
Park
S H
SH
Lee
T H
TH
Nahm
B H
BH
Chung
Y H
YH
Seo
K H
KH
Kim
H Y
HY
D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012478:Salmonella Food Poisoning / Q000175:diagnosis / Q000382:microbiology; D019779:Salmonella enterica / Q000145:classification* / Q000302:isolation & purification*; D012486:Salmonella typhimurium / Q000145:classification / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity; D037521:Virulence Factors / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial; D037521:Virulence Factors