Isolation, Extraction, and Characterization of Verotoxin-producing Escherichia coli O157:H7 from Diarrheal Stool Samples

Muhammad Musthafa Poyil, Ponmurugan Karuppiah, Suresh S. S. Raja, P. Sasikumar
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引用次数: 1

Abstract

Background: Escherichia coli O157:H7 infection causes hemorrhagic colitis and is diagnosed based on symptoms such as cramps, stomach pain, and watery diarrhea. Shiga-like toxins (Verotoxin) produced by Escherichia coli O157:H7 damages endothelial cells of both kidney and brain, causing renal dysfunction and neurological problems. Methods: The present study focuses on identifying the prevalence of Verotoxin-producing Escherichia coli O157:H7 among diarrheal inpatients at Erode Government Hospital, India, and its antibiogram. Further, the Verotoxins were characterized by using SDS-PAGE analysis. A total of 123 samples were collected both from diarrheal stools, and strains from 37 samples (43.02 %) were found to have the presence of E. coli.  The organisms were identified based on their colony morphology on various media, cell morphology, and biochemical tests. The Shiga-like toxin production was identified by non-fermentation of sorbitol on SMAC agar plates. Confirmation of Shiga-like toxin was performed using agglutination assay.  Results: In total, 12 isolates showed agglutination and these isolates were confirmed to be E. coli O157:H7. The molecular weight of the Verotoxin was found to be between 20 and 29 kD. The antibiogram profile of the four isolated strains against 10 standard antibiotics was determined. Conclusion: The results of this study show the occurrence of drug resistance on hemorrhagic colitis causing E. coli O157:H7.
腹泻粪便样品中产维罗毒素大肠杆菌O157:H7的分离、提取和鉴定
背景:大肠杆菌O157:H7感染引起出血性结肠炎,可根据痉挛、胃痛和水样腹泻等症状进行诊断。由大肠杆菌O157:H7产生的志贺样毒素(Verotoxin)损害肾和脑内皮细胞,引起肾功能障碍和神经系统问题。方法:本研究的重点是确定产维罗毒素大肠杆菌O157:H7在印度罗德政府医院腹泻住院患者中的流行情况及其抗生素谱。此外,使用SDS-PAGE分析对Verotoxins进行了表征。从腹泻便中采集的123份样本中,检出大肠杆菌37份(43.02%)。这些微生物是根据它们在不同培养基上的菌落形态、细胞形态和生化试验来鉴定的。通过在SMAC琼脂平板上不发酵山梨醇,鉴定了志贺样毒素的产生。用凝集试验证实志贺样毒素的存在。结果:12株出现凝集现象,经鉴定为大肠杆菌O157:H7。Verotoxin的分子量在20 ~ 29kd之间。测定了4株分离菌株对10种标准抗生素的抗菌谱。结论:本研究结果表明O157:H7大肠杆菌引起的出血性结肠炎存在耐药性。
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