{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T12:04:45Z","timestamp":1789560285001,"version":"build-2803163510"},"reference-count":30,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2011,2,1]],"date-time":"2011-02-01T00:00:00Z","timestamp":1296518400000},"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 Vaccine Immunol"],"published-print":{"date-parts":[[2011,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Infections with methicillin-resistant<jats:italic>Staphylococcus aureus<\/jats:italic>(MRSA) and methicillin-resistant coagulase-negative<jats:italic>Staphylococcus<\/jats:italic>(MR-CNS) are a serious problem in hospitals because these bacteria produce penicillin-binding protein 2\u2032 (PBP2\u2032 or PBP2a), which shows low affinity to \u03b2-lactam antibiotics. Furthermore, the bacteria show resistance to a variety of antibiotics. Identification of these pathogens has been carried out mainly by the oxacillin susceptibility test, which takes several days to produce a reliable result. We developed a simple immunochromatographic test that enabled the detection of PBP2\u2032 within about 20 min. Anti-PBP2\u2032 monoclonal antibodies were produced by a hybridoma of recombinant PBP2\u2032 (rPBP2\u2032)-immunized mouse spleen cells and myeloma cells. The monoclonal antibodies reacted only with PBP2\u2032 of whole-cell extracts and showed no detectable cross-reactivity with extracts from other bacterial species tested so far. One of the monoclonal antibodies was conjugated with gold colloid particles, which react with PBP2\u2032, and another antibody was immobilized on a nitrocellulose membrane, which captures the PBP2\u2032-gold colloid particle complex on a nitrocellulose strip. This strip was able to detect 1.0 ng of rPBP2\u2032 or 2.8 \u00d7 10<jats:sup>5<\/jats:sup>to 1.7 \u00d7 10<jats:sup>7<\/jats:sup>CFU of MRSA cells. The cross-reactivity test using 15 bacterial species and a<jats:italic>Candida albicans<\/jats:italic>strain showed no detectable false-positive results. The accuracy of this method in the detection of MRSA and MR-CNS appeared to be 100%, compared with the results obtained by PCR amplification of the PBP2\u2032 gene,<jats:italic>mecA<\/jats:italic>. This newly developed immunochromatographic test can be used for simple and accurate detection of PBP2\u2032-producing cells in clinical laboratories.<\/jats:p>","DOI":"10.1128\/cvi.00252-10","type":"journal-article","created":{"date-parts":[[2010,12,23]],"date-time":"2010-12-23T03:29:26Z","timestamp":1293074966000},"page":"248-253","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":20,"title":["Development of an Immunochromatographic Strip for Simple Detection of Penicillin-Binding Protein 2\u2032"],"prefix":"10.1128","volume":"18","author":[{"given":"Hidehito","family":"Matsui","sequence":"first","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideaki","family":"Hanaki","sequence":"additional","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Megumi","family":"Inoue","sequence":"additional","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroyuki","family":"Akama","sequence":"additional","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taiji","family":"Nakae","sequence":"additional","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keisuke","family":"Sunakawa","sequence":"additional","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satoshi","family":"Omura","sequence":"additional","affiliation":[{"name":"Laboratory for Antimicrobial Agents, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"},{"name":"Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-Ku, Tokyo 108-8641, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.5.1591-1594.1999"},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"e337","DOI":"10.1371\/journal.pmed.0030337","volume":"3","year":"2006","unstructured":"Chanteau, S., et al. 2006. 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