Laboratory of HealthCare Associated Infection, Centre for Infections, Health Protection Agency, London NW9 5EQ, UK.
HealthCare Associated Infection and Antimicrobial Resistance Department, Centre for Infections, Health Protection Agency, London NW9 5EQ, UK.
J Med Microbiol. 2010 May;59(Pt 5):541-547. doi: 10.1099/jmm.0.015198-0. Epub 2010 Jan 28.
A multiplex PCR is described which detects capsular types K1, K2, K5, K54 and K57, which are those most associated with invasive disease or pathogenicity, a further capsular type (K20), two putative virulence factors (rmpA and wcaG) and the 16S-23S internal transcribed spacer unit of Klebsiella pneumoniae, facilitating identification of this organism. wcaG encodes capsular fucose production and was associated with capsular types K1 and K54, but was also found in strains of other capsular types; 18 of the 543 isolates screened were PCR-positive for this gene. An eight-locus variable number tandem repeat (VNTR) scheme was designed, which provided discrimination at a level similar to that afforded by PFGE among a panel of 36 isolates representing 29 PFGE types. All isolates tested of the virulent K1 clone of CC23, associated with pyogenic liver abscesses, shared the same VNTR profile, which may be helpful in identifying this clone; such isolates were also PCR-positive for allS. These methods provide a rapid means of characterizing and typing isolates of this important agent of community-acquired and nosocomial infection.
描述了一种多重 PCR 方法，可检测与侵袭性疾病或致病性最相关的荚膜型 K1、K2、K5、K54 和 K57，此外还可检测到一种荚膜型（K20）、两种假定的毒力因子（rmpA 和 wcaG）和肺炎克雷伯菌的 16S-23S 内部转录间隔区单元，有助于鉴定该生物体。wcaG 编码荚膜岩藻糖的产生，与荚膜型 K1 和 K54 相关，但也存在于其他荚膜型的菌株中；在筛选的 543 株分离株中，有 18 株对此基因呈 PCR 阳性。设计了一个 8 个基因座可变数串联重复（VNTR）方案，该方案在代表 29 种 PFGE 型的 36 个分离株的小组中提供了类似于 PFGE 的区分水平。所有测试的与化脓性肝脓肿相关的毒力 K1 克隆的 CC23 分离株都具有相同的 VNTR 图谱，这可能有助于识别该克隆；这些分离株也对所有 S 呈 PCR 阳性。这些方法为鉴定这种重要的社区获得性和医院获得性感染病原体的分离株提供了一种快速的特征和分型方法。
