Analysis of virulence factors in Salmonella species

沙门氏菌毒力因子分析

基本信息

批准号：
09041172
负责人：
KURAZONO Hisao
金额：
$ 0.96万
依托单位：
University of Tukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1997
资助国家：
日本
起止时间：
1997 至无数据
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09041172/
关键词：
Salmonella food borne disease Protein toxin Diagmosis Omtibody

项目摘要

At present, over 2,000 serovars of Salmonella species have been recorded with bacterial isolates from various laboratories where the antisera against different serovars of Salmonella are available. The identification of bacterial isolates, however, has been so laborious and well-trained personnel are needed even in the already well-established laboratories. In developing countries, it is hard to have a complete set of biochemical and serological materials for the identification of Salmonella. To solve this problem, we constructed the rapid diagnosis system for the detection of virulent Salmonella species by using the polymerase Chain Reaction (PCR) method.Five hundred sixty seven strains of Salmonella with 58 serovars isolated form clinical specimens and food samples were dedicated to the PCR system. As the stn101 and stn111 primers for enterotoxin gene were used to amplify the complementary target DNA in the samples. It was found that all strains have shown the 260 bp specific DNA fra … More gment in the PCR products as demonstrated by 0.0% agarose gel-electrophoresisWhen PCR were performed by using sin106 and sin112 primers for enteroinvasive gene, the expected 378bp DNA fragment were also found in PCR products of all these Salmonella isolates except for one strain of Salmonella serovar Rissen isolated from the patient with gastroenteritis. These specific primers for enterotoxin and enteroinvasive genes did not show any positive result with the other 224 strains of 30 different bacterial species including enteric bacteria. Confirmation of the PCR products by southern blot hybridization test using synthetic oligonucleotide probes which positions corresponded to stn and invA operons have clearly revealed positive reaction and thus, indicated the specific small DNA fragments in the PCR products obtained from the reaction mixture with stn and sin primers.We also produced anti rabbit-serum against Salmonella enterotoxin and constructed the western blotting system for the detection of it. Less

目前，有2,000种沙门氏菌的血清来自各种经济体的细菌。很难使用一组HE鉴定沙门氏菌。将PCR系统作为STN111和STNTROTOXIN基因耐用，以扩增样品中的靶DNA。通过使用SIN106和SIN112引物进行肠染料基因进行，预期的378BP DNA片段也与肠染色体基因的气体引物一起在使用Southern Blot Hybrot hybridication对Synecter test tess tess tess tess tess tess ccr中发现。对应于STN和Inva操纵子的寡核S清楚地表达了阳性反应，因此来自STH STH SERSO的反应混合物获得的E PCR产物产生了抗菌Ag的抗菌Ag，并构建了它的Western blottem topermem the Western blottem topstem