Ip Margaret, Chau Shirley S L, Chi Fang, Tang Julian, Chan Paul K
Department of Microbiology, Chinese University of Hong Kong, The Prince of Wales Hospital, Hong Kong.
Antimicrob Agents Chemother. 2007 Aug;51(8):2690-700. doi: 10.1128/AAC.00258-07. Epub 2007 Jun 4.
Atypical strains, presumed to be pneumococcus, with ciprofloxacin MICs of > or =4.0 microg/ml and unique sequence variations within the quinolone resistance-determining regions (QRDRs) of the gyrase and topoisomerase genes in comparison with the Streptococcus pneumoniae R6 strain, were examined. These strains were reidentified using phenotypic methods, including detection of optochin susceptibility, bile solubility, and agglutination by serotype-specific antisera, and genotypic methods, including detection of pneumolysin and autolysin genes by PCR, 16S rRNA sequencing, and multilocus sequence typing (MLST). The analysis based on concatenated sequences of the six MLST loci distinguished the "atypical" strains from pneumococci, and these strains clustered closely with S. mitis. However, all these strains and five of nine strains from the viridans streptococcal group possessed one to three gyrA, gyrB, parC, and parE genes whose QRDR sequences clustered with those of S. pneumoniae, providing evidence of horizontal transfer of the QRDRs of the gyrase and topoisomerase genes from pneumococci into viridans streptococci. These genes also conferred fluoroquinolone resistance to viridans streptococci. In addition, the fluoroquinolone resistance determinants of 32 well-characterized Streptococcus mitis and Streptococcus oralis strains from bacteremic patients were also compared. These strains have unique amino acid substitutions in GyrA and ParC that were distinguishable from those in fluoroquinolone-resistant pneumococci and the "atypical" isolates. Both recombinational events and de novo mutations play an important role in the development of fluoroquinolone resistance.
对非典型菌株进行了检测,这些菌株被推测为肺炎球菌,其环丙沙星的最低抑菌浓度(MIC)≥4.0μg/ml,与肺炎链球菌R6菌株相比,其gyrase和topoisomerase基因的喹诺酮耐药决定区(QRDRs)内存在独特的序列变异。使用表型方法对这些菌株进行了重新鉴定,包括检测对奥普托欣的敏感性、胆汁溶解性以及通过血清型特异性抗血清进行凝集试验,还使用了基因型方法,包括通过PCR检测肺炎溶血素和自溶素基因、16S rRNA测序以及多位点序列分型(MLST)。基于六个MLST位点串联序列的分析将“非典型”菌株与肺炎球菌区分开来,这些菌株与缓症链球菌紧密聚类。然而,所有这些菌株以及来自草绿色链球菌群的九株菌株中的五株,都拥有一至三个gyrA、gyrB、parC和parE基因,其QRDR序列与肺炎链球菌的序列聚类,这为gyrase和topoisomerase基因的QRDRs从肺炎球菌水平转移到草绿色链球菌提供了证据。这些基因也赋予了草绿色链球菌对氟喹诺酮的耐药性。此外,还比较了32株来自菌血症患者的特征明确的缓症链球菌和口腔链球菌菌株的氟喹诺酮耐药决定因素。这些菌株在GyrA和ParC中具有独特的氨基酸取代,与耐氟喹诺酮肺炎球菌和“非典型”分离株中的不同。重组事件和从头突变在氟喹诺酮耐药性发展中都起着重要作用。