Pediatric bacteremia and CNS infections associated with klebsiella pneumoniae: molecular genetic characteristics and clinical features

Z. Sadeeva, I. Novikova, Anna V. Lazareva, N. M. Alyabyeva, O. Karaseva, Olga G. Yanushkina, Marina G. Verschinina, A. Fisenko
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Abstract

Klebsiella pneumoniae is one of the most significant and life-threatening pathogen of nosocomial infections. This opportunistic microorganism can cause infections of the bloodstream, respiratory tract, urinary tract, skin and soft tissues, inflammation of meninges of the brain and spinal cord, leading to elevated hospital mortality. The purpose of our study was a retrospective analysis of molecular genetic characteristics of K. pneumoniae isolated from blood and liquor samples as well as to describe clinical features in bacteremia and CNS infections. According to the results of assessed clinical data, K. pneumoniae isolates were selected from 64 children suffered from surgical pathology (congenital heart defects — 30%, abdominal pathology — 39%, severe combined trauma — 12%) and somatic diseases accompanied by antibacterial and/or glucocorticosteroid therapy — 14%. The minimum suppressive concentrations of antibiotics were determined by the broth micro-dilution method. Carbapenemases were detected by real time polymerase chain reaction. Virulence genes and capsule serotypes K1/K2 were assessed by multiplex PCR. Biofilms were grown using flat-bottomed polystyrene plates, followed by coloring, fixation, elution and data detection. The population diversity was assessed by multilocus sequence typing. Bacteremia and CNS infections associated with K. pneumoniae were fatal in 25% of cases. A substantial portion of the isolates demonstrated the phenotype of extremely drug resistance (XDR) — 43%, the phenotype of multidrug resistance (MDR) was shown in 16% of the isolates. The blaCTX-M cephalosporinase gene was found in 85% of the strains. The main determinant of resistance to carbapenems was the blaOXA-48 gene (33%); the blaNDM gene was detected in 9% of strains. The combination of blaOXA-48 and blaNDM was found in 7% of isolates. The study of biofilm production showed that moderate ability to form biofilms was shown in 61%, strong — 21%, and weak — 15% isolates. Two isolates (3%) did not form biofilms. The virulence genes entB and mrkD were detected in 100% of isolates, ybtS — in 78%. The iutA gene was found in 18% of the strains. Two isolates showed the presence of the kfu gene. Seven isolates belonged to the K2 serotype. 27 different genotypes were found in K. pneumoniae isolates examined. The most common were: ST307 — 21%, ST395 — 12%, ST48 — 7%, ST39 — 6% and ST29 — 6%. Infections of the bloodstream and central nervous system associated with K. pneumoniae have great importance in clinical practice. This microorganism is able to long persist on biotic and abiotic surfaces, has a wide natural and acquired resistance to antibiotics.
与肺炎克雷伯菌相关的小儿菌血症和中枢神经系统感染:分子遗传特征和临床特点
肺炎克雷伯氏菌是最重要、最危及生命的院内感染病原体之一。这种机会性微生物可引起血液、呼吸道、泌尿道、皮肤和软组织感染,以及脑膜和脊髓炎症,导致住院死亡率升高。我们的研究旨在对从血液和液体样本中分离出的肺炎克雷伯菌的分子遗传特征进行回顾性分析,并描述菌血症和中枢神经系统感染的临床特征。根据临床数据评估结果,肺炎克雷伯菌分离株选自64名患有外科病变(先天性心脏缺陷-30%、腹部病变-39%、严重合并创伤-12%)和伴有抗菌和/或糖皮质激素治疗的躯体疾病-14%的儿童。抗生素的最低抑制浓度是通过肉汤微稀释法测定的。碳青霉烯酶通过实时聚合酶链反应进行检测。病毒基因和胶囊血清型 K1/K2 通过多重 PCR 进行评估。使用平底聚苯乙烯板培养生物膜,然后进行着色、固定、洗脱和数据检测。通过多焦点序列分型评估种群多样性。在 25% 的病例中,肺炎克氏菌引起的菌血症和中枢神经系统感染是致命的。43%的分离菌株表现出极度耐药(XDR)表型,16%的分离菌株表现出多重耐药(MDR)表型。在 85% 的菌株中发现了 blaCTX-M 头孢菌素酶基因。对碳青霉烯类耐药的主要决定因素是 blaOXA-48 基因(33%);9% 的菌株检测到 blaNDM 基因。在 7% 的分离株中发现了 blaOXA-48 和 blaNDM 基因的组合。对生物膜生成情况的研究表明,61%的分离株具有中等的生物膜生成能力,21%的分离株具有较强的生物膜生成能力,15%的分离株具有较弱的生物膜生成能力。两个分离株(3%)没有形成生物膜。在 100%的分离物中检测到了毒力基因 entB 和 mrkD,在 78% 的分离物中检测到了 ybtS。在 18% 的菌株中发现了 iutA 基因。两个分离株显示存在 kfu 基因。7 个分离株属于 K2 血清型。在检查的肺炎克雷伯菌分离株中发现了 27 种不同的基因型。最常见的是ST307 - 21%、ST395 - 12%、ST48 - 7%、ST39 - 6%和ST29 - 6%。肺炎克雷伯菌引起的血流和中枢神经系统感染在临床实践中具有重要意义。这种微生物能在生物和非生物表面长期存活,对抗生素具有广泛的天然和后天耐药性。
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