Kim H J, Park S H, Lee T H, Nahm B H, Chung Y H, Seo K H, Kim H Y
Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University, Suwon, 449-701, Korea.
J Food Prot. 2006 Jul;69(7):1653-61. doi: 10.4315/0362-028x-69.7.1653.
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.
肠炎沙门氏菌鼠伤寒血清型是全球主要的食源性病原体。到目前为止,尚未开发出用于检测和鉴定鼠伤寒血清型的特定靶基因。为了确定鼠伤寒血清型的特异性探针,利用GenBank中的BLAST(基本局部比对搜索工具)程序选择了预期独特的鼠伤寒血清型LT2基因。通过BLAST将所选基因与各种沙门氏菌血清型的11个基因组序列进行比较。在这些所选基因中,有10个预计对鼠伤寒血清型具有特异性,并且与沙门氏菌致病岛的毒力因子基因或沙门氏菌的O和H抗原基因无关。构建了针对这10个所选基因的引物,并用各种沙门氏菌和非沙门氏菌菌株的基因组DNA对PCR进行评估,以特异性鉴定肠炎沙门氏菌鼠伤寒血清型。在这10个基因的所有引物组中,STM4497对鼠伤寒血清型显示出最高程度的特异性。在本研究中,基于沙门氏菌血清型之间的基因组序列比较,开发了一种针对肠炎沙门氏菌鼠伤寒血清型的特异性引物组,并通过PCR进行了验证。这种通过比较基因组学选择靶基因或序列的方法可应用于微生物的特异性检测。