{"title":"非放射性寡核苷酸探针用于临床产肠毒素大肠杆菌分离株的检测","authors":"A Kumar , M Contrepois , P Tchen , J Cohen","doi":"10.1016/0769-2609(88)90023-3","DOIUrl":null,"url":null,"abstract":"<div><p>Oligonucleotides (32 or 34 mer) corresponding to enterotoxigenic <em>Escherichia coli</em> ST<sub>Ia</sub> (ST-P) toxin were tailed with Bio-11-dUTP using terminal deoxynucleotidyl transferase. Plasmids from clinica <em>E.Coli</em> isolates were prepared by modified rapid alkaline lysis procedure and dot-spotted. Biotinylated oligonucleotide probes were hybridized to detect the St<sub>Ia</sub> toxin gene. The results were in agreement with that obtained by radioactive oligonucleotide probes. Of 135 clinical isolates (sampled from 6 different regions of France), only 7 (5.2 %) were found to be ST<sub>Ia</sub><sup>+</sup>. These 7 isolates were the only ones to be found positive for the K99 adhesive pili antigen. Both the proves were specific to the ST<sub>Ia</sub> toxin gene and failed to detect the closely related ST<sub>Ib</sub> (ST-H) toxin gene. Possibilities of their wide usage in clinical labs are discussed.</p></div>","PeriodicalId":77666,"journal":{"name":"Annales de l'Institut Pasteur. Microbiology","volume":"139 3","pages":"Pages 315-323"},"PeriodicalIF":0.0000,"publicationDate":"1988-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0769-2609(88)90023-3","citationCount":"2","resultStr":"{\"title\":\"Non-radioactive oligonucleotide probe for detection of clinical enterotoxigenic Escherichia Coli isolates of bovine origin\",\"authors\":\"A Kumar , M Contrepois , P Tchen , J Cohen\",\"doi\":\"10.1016/0769-2609(88)90023-3\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Oligonucleotides (32 or 34 mer) corresponding to enterotoxigenic <em>Escherichia coli</em> ST<sub>Ia</sub> (ST-P) toxin were tailed with Bio-11-dUTP using terminal deoxynucleotidyl transferase. Plasmids from clinica <em>E.Coli</em> isolates were prepared by modified rapid alkaline lysis procedure and dot-spotted. Biotinylated oligonucleotide probes were hybridized to detect the St<sub>Ia</sub> toxin gene. The results were in agreement with that obtained by radioactive oligonucleotide probes. Of 135 clinical isolates (sampled from 6 different regions of France), only 7 (5.2 %) were found to be ST<sub>Ia</sub><sup>+</sup>. These 7 isolates were the only ones to be found positive for the K99 adhesive pili antigen. Both the proves were specific to the ST<sub>Ia</sub> toxin gene and failed to detect the closely related ST<sub>Ib</sub> (ST-H) toxin gene. Possibilities of their wide usage in clinical labs are discussed.</p></div>\",\"PeriodicalId\":77666,\"journal\":{\"name\":\"Annales de l'Institut Pasteur. Microbiology\",\"volume\":\"139 3\",\"pages\":\"Pages 315-323\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1988-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0769-2609(88)90023-3\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Annales de l'Institut Pasteur. Microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0769260988900233\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Annales de l'Institut Pasteur. Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0769260988900233","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Non-radioactive oligonucleotide probe for detection of clinical enterotoxigenic Escherichia Coli isolates of bovine origin
Oligonucleotides (32 or 34 mer) corresponding to enterotoxigenic Escherichia coli STIa (ST-P) toxin were tailed with Bio-11-dUTP using terminal deoxynucleotidyl transferase. Plasmids from clinica E.Coli isolates were prepared by modified rapid alkaline lysis procedure and dot-spotted. Biotinylated oligonucleotide probes were hybridized to detect the StIa toxin gene. The results were in agreement with that obtained by radioactive oligonucleotide probes. Of 135 clinical isolates (sampled from 6 different regions of France), only 7 (5.2 %) were found to be STIa+. These 7 isolates were the only ones to be found positive for the K99 adhesive pili antigen. Both the proves were specific to the STIa toxin gene and failed to detect the closely related STIb (ST-H) toxin gene. Possibilities of their wide usage in clinical labs are discussed.
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