Molecular characterization and distribution of cephalosporin resistance determinants in Escherichia coli and Klebsiella pneumoniae isolated from patients attending Kampala International University Teaching Hospital in Bushenyi, Western Uganda

Pub Date : 2021-01-01 DOI:10.1080/20905068.2021.1952821

Herbert Mbyemeire, Kenneth Ssekatawa, C. Kato, E. Wampande

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引用次数: 1

Abstract

ABSTRACT Cephalosporins are the first-line therapy antibiotics used in the treatment of gram-negative bacterial infections. However, high prevalence of cephalosporins resistance in Klebsiella pneumoniae and Escherichia coli has been reported worldwide. Studies conducted in Uganda reported high incidences of cephalosporin resistance (CR). Successive studies at Mulago National Referral Hospital indicated a decline in the resistance levels pointing to the need for regular antibiotic resistance surveillance. Therefore, this study carried out molecular characterization of CR determinants in E. coli and K. pneumoniae isolated from patients attending Kampala International University Teaching Hospital (KIU-TH). A retrospective study using E. coli and K. pnuemoniae samples previously obtained from surgical wounds and urinary tract infections among patients treated at KIU-TH between September 2016 and August 2018 was conducted. Biochemical assays were used to confirm the identity of the samples. Combined disc and boronic acid assays were used to determine the cephalosporine resistance profile of the isolates. Multiplex PCR amplification was used to characterize the extended spectrum beta-lactmase (ESBL) encoding genes. The study revealed that E. coli (130/81.2%) isolates were more predominant than K. pneumoniae (30/18.8%) among the archived samples. K. pneumoniae showed the highest phenotypic resistance with a mean prevalence of 90.6% but comparable to that of E. coli (89.3%). Of the 160 isolates screened, 105 (65.6%) were ESBL producers. Multiplex PCR revealed that the most predominant ESBL encoding gene was bla SHV at a prevalence of 42.0%, followed by bla TEM at 27.3%, bla CTX-M at 22.4% and bla CTX-M-15 at 8.4%. The incidence of phenotypic resistance and distribution of ESBL genes were significantly higher in patients of Ishaka division. Our study reports a high prevalence of cephalosporin-resistant E. coli and K. pnuemoniae isolated from patients attending KIU-TH and highlights the need for routine screening of antimicrobial resistance in health-care facilities so as to guide clinicians on the rational prescription of antibiotics. List of abbreviations: ATCC: American Type Culture Collection; ESBL: extended spectrum beta-lactmases; AmpC: aminopenicillin cephalosporinase; CTX-M: cephotaxime (M-first detected in Munich) hydrolyzing capabilities; CTX-M-U: cephotaxime hydrolyzing capabilities gene Universal primer; SHV: sulfhydryl variables (variant-2); CMY-2: cephamycins (variant-2); TEM: temoneira; ACT-1: AmpC type (variant-1); Bla: Beta lactam; DNA: deoxyribonucleic acid; PCR: Polymerase chain reaction; MNRH: Mulago National Referral Hospital; MRRH: Mbarara Regional Referral Hospital; KRRH: Kabale Regional Referral Hospital (KRRH); KIU-TH: Kampala International University Teaching Hospital; CDL: Central Diagnostic Laboratory; CoVAB: College of Veterinary Medicine, Animal Resources and Biosecurity; CR: cephalosporin resistance

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乌干达西部布申伊坎帕拉国际大学教学医院患者分离的大肠杆菌和肺炎克雷伯菌中头孢菌素耐药性决定簇的分子特征和分布

摘要头孢菌素类抗生素是治疗革兰氏阴性菌感染的一线抗生素。然而，头孢菌素耐药性在肺炎克雷伯菌和大肠杆菌中的高流行率已在世界范围内报道。在乌干达进行的研究报告了头孢菌素耐药性（CR）的高发病率。穆拉戈国家转诊医院的连续研究表明，耐药性水平有所下降，这表明需要定期监测抗生素耐药性。因此，本研究对坎帕拉国际大学教学医院（KIU-TH）患者分离的大肠杆菌和肺炎克雷伯菌中的CR决定簇进行了分子表征。对2016年9月至2018年8月期间在KIU-TH接受治疗的患者中先前从手术伤口和尿路感染中获得的大肠杆菌和假单胞菌样本进行了一项回顾性研究。使用生化分析来确认样品的身份。采用圆盘和硼酸联合测定法测定分离株的头孢菌素耐药性。多重PCR扩增用于表征编码扩展谱β-乳糖酶（ESBL）的基因。研究表明，在存档样本中，大肠杆菌（130/81.2%）分离株比肺炎克雷伯菌（30/18.8%）更占优势。肺炎克雷伯菌表现出最高的表型耐药性，平均患病率为90.6%，但与大肠杆菌（89.3%）相当。在筛选的160个分离株中，105个（65.6%）是ESBL产生者。多重PCR结果显示，ESBL编码基因以bla SHV为主，占42.0%，其次为bla TEM，占27.3%，bla CTX-M，占22.4%，bla CTX-M-15，占8.4%。我们的研究报告了从KIU-TH就诊的患者中分离出的头孢菌素耐药性大肠杆菌和假单胞菌的高流行率，并强调了在卫生保健机构进行常规抗菌药物耐药性筛查的必要性，以指导临床医生合理开用抗生素。缩写列表：ATCC：美国典型培养物保藏中心；ESBL：广谱β-乳糖酶；AmpC：氨基青霉素头孢菌素；CTX-M：cephotaxime（慕尼黑首次检测到M）水解能力；CTX-M-U：头孢他肟水解能力基因通用引物；SHV：巯基变量（变量-2）；CMY-2：头孢霉素（变种-2）；TEM：temoneira；ACT-1：AmpC型（变体-1）；Bla：β-内酰胺；脱氧核糖核酸；聚合酶链式反应；MNRH：穆拉戈国家转诊医院；MRRH：姆巴拉拉地区转诊医院；KRRH：卡巴莱地区转诊医院（KRRH）；KIU-TH：坎帕拉国际大学教学医院；CDL：中央诊断实验室；CoVAB：兽医、动物资源和生物安全学院；CR：头孢菌素耐药性

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