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Rapid detection of the fluoroquinolone resistance‐associated ParC mutation in Neisseria gonorrhoeae using TaqMan probes
Author(s) -
SHIGEMURA KATSUMI,
SHIRAKAWA TOSHIRO,
TANAKA KAZUSHI,
ARAKAWA SOICHI,
GOTOH AKINOBU,
FUJISAWA MASATO
Publication year - 2006
Publication title -
international journal of urology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 67
eISSN - 1442-2042
pISSN - 0919-8172
DOI - 10.1111/j.1442-2042.2006.01264.x
Subject(s) - neisseria gonorrhoeae , taqman , point mutation , serine , polymerase chain reaction , medicine , proline , mutation , microbiology and biotechnology , biology , amino acid , gene , genetics , phosphorylation
Backgrounds: In fluoroquinolone‐resistant Neisseria gonorrhoeae , the amino acid mutations in the fluoroquinolone‐resistant determining region (QRDR) of the parC gene are an important factor. The aim of the present study was to develop a rapid detection method of a serine 88 to proline substitution in parC which we previously showed as having significantly higher fluoroquinolone minimal inhibitory concentrations (MIC) using the TaqMan discrimination system. Methods: We extracted DNA from 90 urine or urethral swab samples obtained from male patients with urethritis caused by N. gonorrhoeae . After DNA extraction, they were subjected to real‐time polymerase chain reaction (PCR) using a TaqMan discrimination system and compared with the results of conventional DNA sequencing. Results: Of the 90 samples, the TaqMan technique result showed 13 samples that were classified as having a serine 88 to proline mutation in parC , and 77 samples that did not have a serine 88 to proline mutation in parC . The classifications of all samples completely corresponded to those determined by conventional DNA sequencing. We also found that N. gonorrhoeae with a serine 88 to proline mutation in parC have a significantly higher MIC to ciprofloxacin than that without a serine 88 to proline mutant in parC . Conclusions: The present genotyping method of real‐time PCR using a TaqMan discrimination system could be applied to the rapid detection of a serine 88 to proline amino acid mutation in parC of N. gonorrhoeae . This point mutation is significant for the determination of fluoruquinolone resistance. This rapid detection system may lead to the prevention of use of noneffective antimicrobial agents and a decrease of resistant strains.