{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T01:49:55Z","timestamp":1762998595748},"reference-count":16,"publisher":"Oxford University Press (OUP)","issue":"12","license":[{"start":{"date-parts":[[2006,12,1]],"date-time":"2006-12-01T00:00:00Z","timestamp":1164931200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2006,12,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Background: For many years, the pathogenic bacterium Neisseria gonorrhoeae, the etiologic agent of gonorrhea, was generally susceptible to penicillin, until the emergence of resistant strains. Well-characterized genetic variations in the penicillin resistance\u2013determining region correlate with decreased susceptibility to penicillin. At least 5 genes (penA, penB, mtrR, ponA, and penC) are involved in the chromosomally mediated resistance to this antibiotic. To date, no development of multiplex PCR assays targeting a range of gonococcal genes and variations as a means of predicting antibiotic resistance has been reported.<\/jats:p>\n               <jats:p>Methods: The aim of this study was to develop a duplex assay using DNA from isolated strains. We describe the development and evaluation on the LightCycler platform of a real-time duplex PCR assay (hybridization probe format) for rapid and specific detection of ponA and penA variations, predicting penicillin susceptibilities.<\/jats:p>\n               <jats:p>Results: The real-time duplex PCR assay successfully detected variations in ponA and penA genes by use of distinct melting temperatures from a total of 120 Neisseria gonorrhoeae isolates. Moreover, the variation profiles obtained with the real-time PCR and the melting analysis showed good correlation with the pattern of penicillin susceptibility generated with classical antibiograms. Nucleotide sequencing data were in complete agreement with multiplex assay results.<\/jats:p>\n               <jats:p>Conclusions: The presented assay is suitable for the detection of chromosomally mediated resistant strains of Neisseria gonorrhoeae in genotyping studies and could be valuable in the effective antimicrobial strategy to gonococci.<\/jats:p>","DOI":"10.1373\/clinchem.2006.075309","type":"journal-article","created":{"date-parts":[[2006,10,20]],"date-time":"2006-10-20T01:30:01Z","timestamp":1161307801000},"page":"2294-2296","source":"Crossref","is-referenced-by-count":17,"title":["A Novel Real-Time Duplex PCR Assay for Detecting penA and ponA Genotypes in Neisseria gonorrhoeae: Comparison with Phenotypes Determined by the E-Test"],"prefix":"10.1093","volume":"52","author":[{"given":"Fre\u0301de\u0301rique","family":"Vernel-Pauillac","sequence":"first","affiliation":[{"name":"Laboratoire de Recherche en Bacte\u0301riologie, Institut Pasteur de Nouvelle-Cale\u0301donie, Nouvelle-Cale\u0301donie"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabrice","family":"Merien","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2006,12,1]]},"reference":[{"key":"2020022801312617900_R1","doi-asserted-by":"crossref","unstructured":"Easmon CSF. The changing pattern of antibiotic resistance of Neisseria gonorrhoeae. Genitourin Med1990;66:55-56.","DOI":"10.1136\/sti.66.2.55"},{"key":"2020022801312617900_R2","doi-asserted-by":"crossref","unstructured":"Herida M, Sednaoui P, Goulet V. Gonorrhoea surveillance system in France: 1986\u20132000. Sex Transm Dis2004;4:209-214.","DOI":"10.1097\/01.OLQ.0000118426.66742.9E"},{"key":"2020022801312617900_R3","doi-asserted-by":"crossref","unstructured":"Ito M, Yasuda M, Yokoi S, Ito S, Takahashi Y, Ishihara S, et al. Remarkable increase in Central Japan in 2001\u20132002 of Neisseria gonorrhoeae isolates with decreased susceptibility to penicillin, tetracycline, oral cephalosporins, and fluoroquinolones. Antimicrob Agents Chemother2004;48:3185-3187.","DOI":"10.1128\/AAC.48.8.3185-3187.2004"},{"key":"2020022801312617900_R4","doi-asserted-by":"crossref","unstructured":"Brannigan JA, Tirodimos IA, Zhang QY, Dowson CG, Spratt BG. Insertion of an extra amino acid is the main cause of the low affinity of penicillin-binding protein 2 in penicillin-resistant strains of Neisseria gonorrhoeae. Mol Microbiol1990;4:913-919.","DOI":"10.1111\/j.1365-2958.1990.tb00664.x"},{"key":"2020022801312617900_R5","doi-asserted-by":"crossref","unstructured":"Dowson CG, Jephcott AE, Gough KR, Spratt BG. Penicillin-binding protein 2 genes of non-\u03b2-lactamase-producing, penicillin-resistant strains of Neisseria gonorrhoeae. Mol Microbiol1989;3:35-41.","DOI":"10.1111\/j.1365-2958.1989.tb00101.x"},{"key":"2020022801312617900_R6","doi-asserted-by":"crossref","unstructured":"Ropp PA, Hu M, Olesky M, Nicholas RA. Mutations in ponA, the gene encoding penicillin-binding protein 1, and a novel locus, penC, are required for high-level chromosomally mediated penicillin resistance in Neisseria gonorrhoeae. Antimicrob Agents Chemother2002;46:769-777.","DOI":"10.1128\/AAC.46.3.769-777.2002"},{"key":"2020022801312617900_R7","doi-asserted-by":"crossref","unstructured":"Paule SM, Trick WE, Tenover FC, Lankford M, Cunningham S, Stosor V, et al. Comparison of PCR assay to culture for surveillance detection of vancomycin-resistant enterococci. J Clin Microbiol2003;41:4805-4807.","DOI":"10.1128\/JCM.41.10.4805-4807.2003"},{"key":"2020022801312617900_R8","doi-asserted-by":"crossref","unstructured":"Lapierre P, Huletsky A, Fortin V, Picard FJ, Roy PH, Ouellette M, et al. Real-time PCR assay for detection of fluoroquinolone resistance associated with grlA mutations in Staphylococcus aureus. J Clin Microbiol2003;41:3246-3251.","DOI":"10.1128\/JCM.41.7.3246-3251.2003"},{"key":"2020022801312617900_R9","doi-asserted-by":"crossref","unstructured":"Stefanelli P, Carattoli A, Neri A, Fazio C, Mastrantonio P. Prediction of decreased susceptibility to penicillin of Neisseria meningitides strains by real-time PCR. J Clin Microbiol2003;41:4666-4670.","DOI":"10.1128\/JCM.41.10.4666-4670.2003"},{"key":"2020022801312617900_R10","doi-asserted-by":"crossref","unstructured":"Vernel-Pauillac F, Falcot V, Whiley D, Merien F. Rapid detection of a chromosomally mediated penicillin resistance-associated ponA mutation in Neisseria gonorrhoeae using a real-time PCR assay. FEMS Microbiol Lett2006;255:66-74.","DOI":"10.1111\/j.1574-6968.2005.00053.x"},{"key":"2020022801312617900_R11","doi-asserted-by":"crossref","unstructured":"Olesky M, Hobbs M, Nicholas RA. Identification and analysis of amino acid mutations in porin IB that mediate intermediate-level resistance to penicillin and tetracycline in Neisseria gonorrhoeae. Antimicrob Agents Chemother2002;46:2811-2820.","DOI":"10.1128\/AAC.46.9.2811-2820.2002"},{"key":"2020022801312617900_R12","doi-asserted-by":"crossref","unstructured":"Tirodimos I, Tzelepi E, Katsougiannopoulos VC. Penicillin-binding protein 2 genes of chromosomally-mediated penicillin-resistant Neisseria gonorrhoeae from Greece: screening for codon Asp-345A. J Antimicrob Chemother1993;32:677-684.","DOI":"10.1093\/jac\/32.5.677"},{"key":"2020022801312617900_R13","doi-asserted-by":"crossref","unstructured":"Veal WL, Nicholas RA, Shafer WM. Overexpression of the MtrC- MtrD- MtrE efflux pump due to an mtrR mutation is required for chromosomally mediated penicillin resistance in Neisseria gonorrhoeae. J Bacteriol2002;184:5619-5624.","DOI":"10.1128\/JB.184.20.5619-5624.2002"},{"key":"2020022801312617900_R14","doi-asserted-by":"crossref","unstructured":"Barratt K, Mackay JF. Improving real-time PCR genotyping assays by asymmetric amplification. 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