Abstract
In order to investigate the pathogenicity and development of differential identification technique in the Yersinia species and other entericbacteria, we isolated 5 strains of Y.pseudotuberculosis from spring water sites in Seoul. The biochemical characteristics of isolated strains revealed that indole, VP($25^{\circ}C$, $37^{\circ}C$), $H_2S$, phenylalanine, lysine, arginine, ornithine, gas from glucose, lactose, sucrose, sorbitol, oxidase and motility($37^{\circ}C$) were all negative and urease, glucose, mannitol, salicin, catalase and motility($25^{\circ}C$) were all positive. To detect the causative agent of pseudotuberculosis(Y.pseudotuberculosis), we carried out a study using a PCR with inv primers complementary to the pathogenic region and found that all strains were positive, this revealed that strains from spring waters were pathogenic. Also 16S rDNA for total 5 strains of Y. pseudotuberculosis were amplified and a stretch of approximately 1,450 nucleotides were sequenced and analyzed. The 16S rDNA nucleotide sequence homologies among Yersinia species ranged 97.5% to 100% and between Y.pseudotuberculosis and other entericbacteria they ranged 93.0% to 95.1%. The Phylogenetic tree generated from the sequence analysis of the 16S rDNA gene showed 3 coherent clusters that could be separated into Y.pseudotuberculsis strains, some Yersinia species strains and other entericbacteria strains.
서울시내 25개 자치구에 산재한 약수터에서 5주의 Yersinia pseudotuberculosis를 분리하여 생화학적 특성, 병원성의 유무 및 16S rDNA 분석을 실시하였다. 분리된 Y. pseudotuberculosis는 모두 병원성 유전자인 inv를 소유하고 있었으며, 16S rDNA를 증폭한 후 염기서열을 분석하여 NCBI Genbank에 등록된 다른 Yersinia 속 및 장내세균 등과 비교해 본 결과 Yersinia 속 등과는 97.5%에서 100%의 높은 상동성(Similarity)을 나타내었고, 다른 장내세균 등과는 93.0%에서 95.1%의 낮은 상동성을 나타내었다. 165 rDNA 염기서열을 기초로 계통수를 작성한 결과 크게 3개의 cluster를 형성하였는데 특히 Y.enterocolitica (Z49830)은 Y.pseudotuberculasis (Z21939)와의 상동성(97.7%)보다 Y.intermedia (X75279)와의 상동성(97.9%)이 더 높게 나타났으며, E. coli (Z83205)는 Proteus vulgaris (AJ233425) 와의 상동성(93.2%)보다 Salmonella enteritidis (U90318)와의 상동성(97.7%)이 더 밀접한 연관성을 나타내었다.
