{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,23]],"date-time":"2025-09-23T14:40:04Z","timestamp":1758638404967},"reference-count":31,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2000,7,1]],"date-time":"2000-07-01T00:00:00Z","timestamp":962409600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Clin Diagn Lab Immunol"],"published-print":{"date-parts":[[2000,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The p36 protein of\n            <jats:italic>Mycoplasma hyopneumoniae<\/jats:italic>\n            is a cytosolic protein carrying species-specific antigenic determinants. Based on the genomic sequence of the reference strain ATCC 25934, primers were designed for PCR amplification of the p36-encoding gene (948 bp). These primers were shown to be specific to\n            <jats:italic>M. hyopneumoniae<\/jats:italic>\n            since no DNA amplicons could be obtained with other mycoplasma species and pathogenic bacteria that commonly colonize the porcine respiratory tract. The amplified p36 gene was subcloned into the pGEX-4T-1 vector to be expressed in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            as a fusion protein with glutathione\n            <jats:italic>S<\/jats:italic>\n            -transferase (GST). The GST-p36 recombinant fusion protein was purified by affinity chromatography and cut by thrombin, and the enriched p36 protein was used to immunize female BALB\/c mice for the production of anti-p36 monoclonal antibodies (MAbs). The polypeptide specificity of the nine MAbs obtained was confirmed by Western immunoblotting with cell lysates prepared from the homologous strain. Cross-reactivity studies of the anti-p36 MAbs towards two other\n            <jats:italic>M. hyopneumoniae<\/jats:italic>\n            reference strains (ATCC 25095 and J strains) and Quebec field strains that had been isolated in culture suggested that these anti-p36 MAbs were directed against a highly conserved epitope, or closely located epitopes, of the p36 protein. No reactivity was demonstrated against other mycoplasma species tested. Clinical signs and lesions suggestive of enzootic pneumonia were reproduced in specific-pathogen-free pigs infected experimentally with a virulent Quebec field strain (IAF-DM9827) of\n            <jats:italic>M. hyopneumoniae<\/jats:italic>\n            . The bacteria could be recovered from lung homogenates of pigs that were killed after the 3-week observation period by both PCR and cultivation procedures. Furthermore, the anti-p36 MAbs permitted effective detection by indirect immunofluorescence of\n            <jats:italic>M. hyopneumoniae<\/jats:italic>\n            in frozen lung sections from experimentally infected pigs. However, attempts to use the recombinant p36 protein as an antigen in an indirect enzyme-linked immunosorbent assay for the detection of antibodies in sera from convalescent pigs showed no correlation with clinical and pathological findings.\n          <\/jats:p>","DOI":"10.1128\/cdli.7.4.528-535.2000","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:03:53Z","timestamp":1027764233000},"page":"528-535","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["Species-Specific Monoclonal Antibodies to\n            <i>Escherichia coli<\/i>\n            -Expressed p36 Cytosolic Protein of\n            <i>Mycoplasma hyopneumoniae<\/i>"],"prefix":"10.1128","volume":"7","author":[{"given":"J.","family":"Caron","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Universite\u0301 du Que\u0301bec, Laval, Que\u0301bec H7V 1B7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N.","family":"Sawyer","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Universite\u0301 du Que\u0301bec, Laval, Que\u0301bec H7V 1B7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B. Ben Abdel","family":"Moumen","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Universite\u0301 du Que\u0301bec, Laval, Que\u0301bec H7V 1B7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K. Cheikh Saad","family":"Bouh","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Universite\u0301 du Que\u0301bec, Laval, Que\u0301bec H7V 1B7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S.","family":"Dea","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Universite\u0301 du Que\u0301bec, Laval, Que\u0301bec H7V 1B7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"1349","article-title":"Diagnosis of mycoplasmal pneumonia of swine: sequential study by direct immunofluorescence.","volume":"45","author":"Amanfu W.","year":"1984","unstructured":"Amanfu W. Weng C. N. Ross R. F. Barnes H. J. Diagnosis of mycoplasmal pneumonia of swine: sequential study by direct immunofluorescence. Am. J. Vet. 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The Mycoplasma hyopneumoniae 74.5kD antigen elicits neutralizing antibodies and shares sequence similarity with heat-shock proteins. Vaccine 89 1989 265 269","journal-title":"Vaccine"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.4.1390-1396.2000"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-70-7-1725"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1177\/104063879100300401"},{"key":"e_1_3_2_8_2","unstructured":"Feenstra\nA. A.\nS\u00f6rensen\nV.\nFriis\nN. F.\nJensen\nN. 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