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使用基于培养和酶联免疫吸附测定的毒素检测方法检测一种被忽视的病原体:来自三级医疗机构的单中心研究。

Use of culture- and ELISA-based toxin assay for detecting , a neglected pathogen: A single-center study from a tertiary care setting.

作者信息

Lall Sujata, Nataraj Gita, Mehta Preeti

机构信息

Department of Microbiology, Seth GSMC and KEMH, Mumbai, Maharashtra, India.

出版信息

J Lab Physicians. 2017 Oct-Dec;9(4):254-259. doi: 10.4103/JLP.JLP_157_16.

Abstract

INTRODUCTION

Clostridium difficile is a Gram-positive spore-bearing anaerobic bacillus increasingly associated with both community- and hospital-acquired colitis and diarrhea. It is the most common identifiable bacterial cause of healthcare-associated diarrhea associated with antibiotic use and one of the most common anaerobic infections. The diagnosis of C. difficile infection includes detection of toxin A/B in stool specimens by direct enzyme immunoassay, culture of pathogen from the stool specimens using a selective agar Cycloserine-Cefoxitin fructose agar (CCFA), tissue culture assay, and detection of glutamate dehydrogenase an enzyme produced by C. difficile. With few reports from India on this disease, the present study was planned to throw more light on the prevalence and utility of laboratory diagnostic methods for C. difficile-associated diarrhea (CDAD).

MATERIAL AND METHODS

After taking approval from the Ethics Committee, 150 patients with antibiotic-associated diarrhea were taken as a study group and fifty patients with exposure to antibiotics but who did not develop diarrhea were taken as controls. Stool specimen was processed for both culture on CCFA and toxin detection by IVD Tox A + B ELISA.

RESULTS

Only four specimens were culture positive, whereas 13 were ELISA positive. All culture-positive isolates were toxigenic. C. difficile was neither isolated nor its toxin detected in the control group. Culture- and toxin-based assays may not detect all cases of CDAD.

CONCLUSION

Based on the results of the present study, culture does not provide any additional yield over toxin assay. Better diagnostic modalities would be required to prove CDAD.

摘要

引言

艰难梭菌是一种革兰氏阳性、产芽孢的厌氧杆菌,越来越多地与社区获得性和医院获得性结肠炎及腹泻相关。它是与抗生素使用相关的医疗保健相关腹泻最常见的可识别细菌病因,也是最常见的厌氧菌感染之一。艰难梭菌感染的诊断包括通过直接酶免疫测定法检测粪便标本中的毒素A/B、使用选择性琼脂环丝氨酸-头孢西丁果糖琼脂(CCFA)从粪便标本中培养病原体、组织培养测定法以及检测艰难梭菌产生的一种酶谷氨酸脱氢酶。由于印度关于这种疾病的报道很少,本研究旨在进一步阐明艰难梭菌相关性腹泻(CDAD)的患病率及实验室诊断方法的实用性。

材料与方法

经伦理委员会批准,将150例抗生素相关性腹泻患者作为研究组,50例接触过抗生素但未发生腹泻的患者作为对照组。粪便标本分别进行CCFA培养和IVD Tox A + B ELISA毒素检测。

结果

仅4份标本培养阳性,而ELISA阳性的有13份。所有培养阳性的分离株均产毒素。对照组未分离出艰难梭菌,也未检测到其毒素。基于培养和毒素的检测方法可能无法检测出所有CDAD病例。

结论

根据本研究结果,培养法相对于毒素检测法并无额外收获。需要更好的诊断方法来确诊CDAD。

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