Carter P E, Abadi F J, Yakubu D E, Pennington T H
Department of Medical Microbiology, University of Aberdeen Medical School, United Kingdom.
Antimicrob Agents Chemother. 1994 Jun;38(6):1256-61. doi: 10.1128/AAC.38.6.1256.
Primers were designed to amplify the rpoB gene of Neisseria meningitidis. The region of the gene amplified covered clusters I and II of the rifampin resistance (Rifr) mutation sites identified in Escherichia coli. DNAs from six Rifr isolates and 21 rifampin-susceptible isolates from the United Kingdom representing a number of serogroups were amplified and sequenced. All six Rifr isolates had identical DNA sequences and the same amino acid change, a His to an Asn change at position 35 (H35N). This His residue is equivalent to the His residue at position 526 in E. coli, one of the known Rifr mutation sites. DNAs from an additional six Rifr mutations generated in vitro were amplified and sequenced. Three had H35Y changes, one had an H35R change, one had an H35N change and one had an S40F change. The predominance of mutations at the His residue at position 35 in Rifr N. meningitidis isolates suggests that it plays a critical role in the selection of antibiotic-resistant variants. All six Rifr isolates belonged to the same clonal group when analyzed by restriction enzyme analysis and pulsed-field gel electrophoresis. These data suggest that a single clone of Rifr N. meningitidis is present and widespread throughout the United Kingdom.
设计引物以扩增脑膜炎奈瑟菌的rpoB基因。扩增的基因区域覆盖了在大肠杆菌中鉴定出的利福平耐药(Rifr)突变位点的I群和II群。对来自英国的代表多个血清群的6株利福平耐药分离株和21株利福平敏感分离株的DNA进行了扩增和测序。所有6株利福平耐药分离株具有相同的DNA序列和相同的氨基酸变化,即第35位的组氨酸(His)变为天冬酰胺(Asn)(H35N)。该组氨酸残基等同于大肠杆菌中第526位的组氨酸残基,这是已知的利福平耐药突变位点之一。对另外6个体外产生的利福平耐药突变的DNA进行了扩增和测序。3个有H35Y变化,1个有H35R变化,1个有H35N变化,1个有S40F变化。脑膜炎奈瑟菌利福平耐药分离株中第35位组氨酸残基处突变的优势表明,它在抗生素耐药变异体的选择中起关键作用。通过限制性酶切分析和脉冲场凝胶电泳分析,所有6株利福平耐药分离株属于同一克隆群。这些数据表明,在英国存在单一克隆的脑膜炎奈瑟菌利福平耐药株且广泛传播。