Early Detection of Drug Resistant Enterobacteriaceae in Urinary Tract Infections using Chromogenic Agar Medium in a Tertiary Care Hospital, Kakinada, Andhra Pradesh, India

N. Reddi, Sobharani Sanapala, Radhika Budumuru
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Abstract

Introduction: The irrational and inappropriate use of beta lactam antimicrobial drugs has led to the advent of Extended Spectrum Beta-Lactamase (ESBL) resistant strains. ESBL producing Enterobacteriaceae strains are frequent causative agents both in community and in acquired nosocomial infections and Urinary Tract Infections (UTI). The phenotypic confirmatory tests rarely identify all ESBLs. Chrom ID (Chromogenic identification Media) ESBL – Bx (bioMerieux) is a completely new and innovative chromogenic medium designed specifically for the screening of ESBL producing Enterobacteria directly from urine samples. It is a ready to use selective media which is sensitive and specific for rapid and presumptive identification of ESBL producing Enterobacteriaceae. Aim: Early detection of ESBL producing Enterobacteriaceae directly from urine samples on chromogenic medium (Chrom ID- ESBL- Bx) and confirmation of ESBL producing Enterobacteria using Disc Potentiation Test (DPT). Materials and Methods: The present cross-sectional study was conducted in the Department of Microbiology, Rangaraya Medical College, Kakinada, Andhra Pradesh, India from November 2019 to March 2020 (five months duration). The study was done on 70 urine samples from patients with UTI. All samples were subjected to wet mount, inoculated directly for culture on Chrom ID ESBL- Bx agar and MacConkey agar. Antibiotic Susceptibility testing of ceftazidime and cefotaxime was done by Kirby-Bauer disc diffusion method and conformation of ESBL production by DPT using Clinical and Laboratory Standards Institute (CLSI) method. The Statistical Package for the Social Sciences (SPSS) Statistical package version (18.0) was used. Results: A total of 56 (80%) isolates were obtained from 70 urine samples, out of them 28 (50%) were Escherichia coli, 21 (37.5%) were Klebsiella spp., 7 (12.5%) were Proteus spp., 23 (82.14%) isolates of Escherichia coli, 15 (71.43%) of Klebsiella spp., 6 (85.71%) of Proteus spp., isolated were screened positive using Chrom ID ESBL-Bx agar. About 44 (78.57%) of total Enterobacteria (56) were screened for ESBL production. 20 (86.96%) of Escherichia coli, 11 (73.33%) of Klebsiella spp., and 5 (83.33%) of Proteus spp., that were screened positive using Chrom ID ESBL agar were confirmed (by DPT) as ESBL producers and 2 (16.6%) of total (12) isolates that were screened negative by Chrom ID ESBL agar were confirmed as ESBL producers when screened and confirmed by DPT. So sensitivity and specificity CHRO Magar was 94.73% and 55.5%. Conclusion: ESBL continues to become a serious public health threat. Results from present study showed that CHROMagar ESBL has a high sensitivity and a convenient method for making provisional diagnosis of drug resistant Enterobacterial infections in 24 hours. Chrom ID ESBL- Bx agar medium allows easy differentiation of different bacteria based on colony colouration.
用显色琼脂培养基在印度安得拉邦Kakinada三级医院早期检测尿路感染中的耐药肠杆菌科
引言:β -内酰胺类抗菌药物的不合理和不适当使用导致了扩展谱β -内酰胺酶(ESBL)耐药菌株的出现。产生ESBL的肠杆菌科菌株是社区和获得性医院感染和尿路感染(UTI)的常见病原体。表型确认试验很少能识别所有esbl。Chrom ID(显色鉴定培养基)ESBL - Bx (bioMerieux)是一种全新的创新显色培养基,专为直接从尿液样本中筛选产生ESBL肠杆菌而设计。它是一种敏感和特异的选择性培养基,可用于快速和推定鉴定产生ESBL的肠杆菌科。目的:用显色培养基(Chrom ID- ESBL- Bx)检测尿样中直接产肠杆菌科ESBL,并采用圆盘增强试验(DPT)确认ESBL产肠杆菌。材料与方法:本横断面研究于2019年11月至2020年3月在印度安得拉邦Kakinada市Rangaraya医学院微生物学系进行(为期5个月)。这项研究是对来自尿路感染患者的70份尿液样本进行的。所有样品均湿载,直接接种于Chrom ID ESBL- Bx琼脂和MacConkey琼脂上培养。采用Kirby-Bauer盘片扩散法对头孢他啶和头孢噻肟进行药敏试验,采用临床与实验室标准协会(CLSI)方法对DPT制备的ESBL进行构象检测。采用SPSS (Statistical Package for The Social Sciences)统计软件包18.0版本。结果:70份尿样共分离到56株(80%),其中大肠埃希菌28株(50%),克雷伯氏菌21株(37.5%),变形杆菌7株(12.5%),大肠埃希菌23株(82.14%),克雷伯氏菌15株(71.43%),变形杆菌6株(85.71%),用Chrom ID esblb - bx琼脂筛选阳性。在56株肠杆菌中筛选到44株(78.57%)产生ESBL。用Chrom ID ESBL琼脂筛选阳性的大肠埃希菌20株(86.96%)、克雷伯氏菌11株(73.33%)、变形杆菌5株(83.33%)被DPT证实为ESBL产生菌,经DPT筛选确认为阴性的12株中有2株(16.6%)为ESBL产生菌。因此,CHRO Magar的敏感性和特异性分别为94.73%和55.5%。结论:ESBL继续成为严重的公共卫生威胁。本研究结果表明,CHROMagar ESBL具有较高的敏感性,是一种方便的24小时耐药肠杆菌感染临时诊断方法。Chrom ID ESBL- Bx琼脂培养基可以根据菌落颜色轻松区分不同的细菌。
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