El Fertas-Aissani R, Messai Y, Alouache S, Bakour R
Laboratoire de biologie cellulaire et moléculaire, faculté des sciences biologiques, université des sciences et de la technologie Houari Boumediene, BP 32 El-Alia, Bab Ezzouar, 16111 Alger, Algeria.
Pathol Biol (Paris). 2013 Oct;61(5):209-16. doi: 10.1016/j.patbio.2012.10.004. Epub 2012 Dec 5.
To detect virulence factors in 54 Klebsiella pneumoniae isolates from different clinical specimens: urine (26), blood (11), pus (11), lung (four), cerebrospinal fluid (one) and ascitic fluid (one).
PCR was used to investigate virulence genes encoding adhesins (fimH-1, mrkD, kpn, ycfM), siderophores (entB: enterobactin, iutA: aerobactin, irp-1, irp-2, ybtS, fyuA: yersiniabactin, iroN: catechols receptor), protectines or invasins (rmpA, magA, traT) and toxins (hlyA, cnf-1). The serum resistance, capsule and hypermucoviscosity, and ability to form biofilm and produce siderophores were sought by phenotypic assays. The in vivo virulence was assessed in mice infected by intraperitoneal way. Antimicrobial susceptibility was tested by diffusion method.
The most common virulence genes were fimH-1 (100%), mrkD (96.3%), ycfM (96.3%), and entB (100%). kpn and yersiniabactin genes were found at medium rates of 63% and 46.3% and at lower prevalence, were genes traT (1.8%), iroN (3.7%), iutA (5.5%) and rmpA (3.7%). magA, hlyA and cnf-1 genes were not detected. The capsule, serum resistance, biofilm formation, mannose-sensitive or -resistant haemagglutination and hypermucoviscosity were observed in 100%, 92.6%, 88.8%, 94.4%, 68.5% and 9.2% of isolates, respectively. The prevalence of siderophores was consistent with that of genotypic detection. The LD50 in mice was very low (<10(2) CFUs) for isolates with the most virulence factors. A rate of 74.1% of isolates showed a multidrug resistance (MDR) pattern.
The distribution of virulence profiles according to the clinical origin suggests a role of enterobactin in urinary infections and yersiniabactin in the invasiveness. The fimbriae F1 and F3, capsule, enterobactin, serum resistance and biofilm formation, were commonly found in isolates, they seem to be at the basis of classic pathogenicity of K. pneumoniae. The invasiveness enhancers, aerobactin, yersiniabactin, catechols receptor, mucoid factor and hypermucoviscosity, detected concomitantly in some isolates, constitute a threat for vulnerable populations, even more if they are in combination with antibiotic resistance.
检测从不同临床标本中分离出的54株肺炎克雷伯菌的毒力因子,这些标本包括尿液(26份)、血液(11份)、脓液(11份)、肺组织(4份)、脑脊液(1份)和腹水(1份)。
采用聚合酶链反应(PCR)检测编码黏附素(fimH-1、mrkD、kpn、ycfM)、铁载体(entB:肠杆菌素、iutA:气杆菌素、irp-1、irp-2、ybtS、fyuA:耶尔森菌素、iroN:儿茶酚受体)、保护素或侵袭素(rmpA、magA、traT)以及毒素(hlyA、cnf-1)的毒力基因。通过表型分析检测血清抗性、荚膜和高黏液性,以及形成生物膜和产生铁载体的能力。通过腹腔注射感染小鼠来评估体内毒力。采用扩散法检测抗菌药物敏感性。
最常见的毒力基因是fimH-1(100%)、mrkD(96.3%)、ycfM(96.3%)和entB(100%)。kpn和耶尔森菌素基因的检出率中等,分别为63%和46.3%,而traT(1.8%)、iroN(3.7%)、iutA(5.5%)和rmpA(3.7%)基因的检出率较低。未检测到magA、hlyA和cnf-1基因。分别在100%、92.6%、88.8%、94.4%、68.5%和9.2%的分离株中观察到荚膜、血清抗性、生物膜形成、甘露糖敏感或抗性血凝反应以及高黏液性。铁载体的流行情况与基因型检测结果一致。对于具有最多毒力因子的分离株,小鼠的半数致死剂量(LD50)非常低(<10²CFU)。74.1%的分离株表现出多重耐药(MDR)模式。
根据临床来源的毒力谱分布表明,肠杆菌素在尿路感染中起作用,而耶尔森菌素在侵袭性方面起作用。分离株中常见菌毛F1和F3、荚膜、肠杆菌素、血清抗性和生物膜形成,它们似乎是肺炎克雷伯菌经典致病性的基础。在一些分离株中同时检测到的侵袭增强因子气杆菌素、耶尔森菌素、儿茶酚受体、黏液样因子和高黏液性,对易感人群构成威胁,尤其是当它们与抗生素耐药性同时存在时。