{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T15:18:18Z","timestamp":1787239098414,"version":"build-2736575974"},"reference-count":20,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2002,1,1]],"date-time":"2002-01-01T00:00:00Z","timestamp":1009843200000},"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":["J Clin Microbiol"],"published-print":{"date-parts":[[2002,1]]},"abstract":"ABSTRACT<\/jats:title>\n \n A rapid real-time multiplex PCR assay for detecting and differentiating\n Bordetella pertussis<\/jats:italic>\n and\n Bordetella parapertussis<\/jats:italic>\n in nasopharyngeal swabs was developed. This assay (LC-PCR-IS) targets the insertion sequences IS\n 481<\/jats:italic>\n and IS\n 1001<\/jats:italic>\n of\n B. pertussis<\/jats:italic>\n and\n B. parapertussis<\/jats:italic>\n , respectively, and is performed using the LightCycler (Roche Molecular Biochemicals, Indianapolis, Ind.). The analytical sensitivity is less than one organism per reaction. Results for\n Bordetella<\/jats:italic>\n culture and\/or direct fluorescent antibody testing and a second LightCycler PCR assay (target, pertussis toxin gene) were compared to results of the LC-PCR-IS assay for 111 nasopharyngeal swabs submitted for pertussis testing. Of the specimens, 12 were positive (9\n B. pertussis<\/jats:italic>\n and 3\n B. parapertussis<\/jats:italic>\n ) and 68 specimens were negative by all methods. Three other specimens were positive for\n B. pertussis<\/jats:italic>\n by at least two of the methods (including the LC-PCR-IS assay), and another 28 specimens were positive for\n B. pertussis<\/jats:italic>\n by the LC-PCR-IS assay only. No specimens were negative by the LC-PCR-IS assay and positive by the other methods. A conventional PCR method (target, IS\n 481<\/jats:italic>\n ) was also compared to the LC-PCR-IS assay for a different group of nasopharyngeal swab specimens (\n n<\/jats:italic>\n = 96): 44 specimens were positive and 41 specimens were negative for\n B. pertussis<\/jats:italic>\n with both PCR methods. Nine specimens were positive for\n B. pertussis<\/jats:italic>\n by the LC-PCR-IS assay and negative by the conventional PCR assay, and two specimens were positive for\n B. pertussis<\/jats:italic>\n by the conventional PCR assay and negative by the LC-PCR-IS assay. Positivity of the two assays was not significantly different (\n P<\/jats:italic>\n = 0.0654). The insertion sequence IS\n 481<\/jats:italic>\n is also present in\n Bordetella holmesii<\/jats:italic>\n ; specimens containing\n B<\/jats:italic>\n .\n holmesii<\/jats:italic>\n may yield false-positive results. The LC-PCR-IS assay takes approximately 45 min to complete post-nucleic acid extraction, compared to 24 h for the conventional PCR assay previously used in our laboratory. The LC-PCR-IS assay is easier to perform than the conventional PCR assay, and the closed system decreases the chance of contamination. All of these characteristics represent a significant improvement in the detection of\n B<\/jats:italic>\n .\n pertussis<\/jats:italic>\n and\n B<\/jats:italic>\n .\n parapertussis<\/jats:italic>\n in nasopharyngeal specimens.\n <\/jats:p>","DOI":"10.1128\/jcm.40.1.96-100.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T06:03:40Z","timestamp":1027749820000},"page":"96-100","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":77,"title":["Multiplex LightCycler PCR Assay for Detection and Differentiation of\n Bordetella pertussis<\/i>\n and\n Bordetella parapertussis<\/i>\n in Nasopharyngeal Specimens"],"prefix":"10.1128","volume":"40","author":[{"given":"Lynne M.","family":"Sloan","sequence":"first","affiliation":[{"name":"Division of Clinical Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marlene K.","family":"Hopkins","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P. Shawn","family":"Mitchell","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily A.","family":"Vetter","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jon E.","family":"Rosenblatt","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"},{"name":"Division of Infectious Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. Scott","family":"Harmsen","sequence":"additional","affiliation":[{"name":"Section of Biostatistics, Mayo Clinic, Rochester, Minnesota"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franklin R.","family":"Cockerill","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"},{"name":"Division of Infectious Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin","family":"Patel","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"},{"name":"Division of Infectious Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"25","volume":"46","year":"1997","unstructured":"Centers for Disease Control and Prevention. 1997. Case definitions for infectious conditions under public health surveillance. Morb. Mortal. Wkly. Rep.46(RR-10):25.","journal-title":"Morb. Mortal. Wkly. 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