{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T00:22:49Z","timestamp":1765758169624},"reference-count":14,"publisher":"Microbiology Society","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2005,5,1]]},"abstract":"<jats:p>Real-time PCR based on the capsule transfer gene (<jats:italic>ctrA<\/jats:italic>) is a significant aid in the diagnosis of meningococcal infection but fails to detect a high proportion (60 %) of non-groupable strains associated with nasopharyngeal carriage. This study aimed to design a novel real-time (TaqMan) PCR that would detect more strains of meningococci and be suitable for large-scale carriage studies. Primer and probe sequences were based on the meningococcal <jats:italic>porA<\/jats:italic> gene and designed specifically to exclude the highly related <jats:italic>porA<\/jats:italic> pseudogene in <jats:italic>Neisseria gonorrhoeae<\/jats:italic>. The specificity of the assay was confirmed by testing strains of <jats:italic>N. gonorrhoeae<\/jats:italic> known to contain the <jats:italic>porA<\/jats:italic> pseudogene together with commensal strains of <jats:italic>Neisseria lactamica<\/jats:italic> and <jats:italic>Neisseria sicca<\/jats:italic>. None of these was detected in the assay. <jats:italic>Neisseria meningitidis<\/jats:italic> strains representing a wide range of serogroups together with non-groupable strains isolated from the nasopharynx were tested by <jats:italic>ctrA<\/jats:italic> assay and the novel <jats:italic>porA<\/jats:italic>-based TaqMan PCR. All carriage strains were detected by the <jats:italic>porA<\/jats:italic>-based assay including four that gave weak or no reaction with the <jats:italic>ctrA<\/jats:italic> assay. Comparison of <jats:italic>ctrA<\/jats:italic> and <jats:italic>porA<\/jats:italic> assays on 71 throat swabs obtained from university students showed that the <jats:italic>porA<\/jats:italic> assay detected meningococcal DNA in all samples that were <jats:italic>ctrA<\/jats:italic> positive plus three that were <jats:italic>ctrA<\/jats:italic> negative but culture positive. This novel <jats:italic>porA<\/jats:italic>-based TaqMan assay provides a highly specific method for detecting meningococcal DNA that is more sensitive than the <jats:italic>ctrA<\/jats:italic> assay for detecting meningococcal carriage and is particularly suitable for carriage studies where non-groupable strains and other <jats:italic>Neisseria<\/jats:italic> are present.<\/jats:p>","DOI":"10.1099\/jmm.0.45847-0","type":"journal-article","created":{"date-parts":[[2005,4,11]],"date-time":"2005-04-11T23:38:02Z","timestamp":1113262682000},"page":"463-466","source":"Crossref","is-referenced-by-count":34,"title":["A novel porA-based real-time PCR for detection of meningococcal carriage"],"prefix":"10.1099","volume":"54","author":[{"given":"J Zoe","family":"Jordens","sequence":"first","affiliation":[{"name":"Southampton Public Health Laboratory1 and Molecular Microbiology & Infection, Division of Infection, Inflammation & Repair,2 University of Southampton Medical School, Southampton General Hospital, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John E","family":"Heckels","sequence":"additional","affiliation":[{"name":"Southampton Public Health Laboratory1 and Molecular Microbiology & Infection, Division of Infection, Inflammation & Repair,2 University of Southampton Medical School, Southampton General Hospital, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"H1-8","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J Mol Biol"},{"key":"H2-8","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1111\/j.1365-2958.1989.tb01802.x","article-title":"The class 1 outer membrane protein of Neisseria meningitidis : gene sequence and structural and immunological similarities to gonococcal 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