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免疫斑点印迹法检测鼠伤寒沙门氏菌中霍乱相关肠毒素抗原的评估。

Evaluation of immuno-dot-blot assay for detection of cholera-related enterotoxin antigen in Salmonella typhimurium.

作者信息

Panigrahi D, Burks M, Hariharan H, Finkelstein R A

出版信息

J Clin Microbiol. 1987 Apr;25(4):702-5. doi: 10.1128/jcm.25.4.702-705.1987.

Abstract

Twenty-five strains of Salmonella typhimurium isolated in India were examined for the presence of cholera/coli-related enterotoxin antigen by a previously described latex particle agglutination test and by a newly developed immuno-dot-blot test using immunopurified goat antibody against the cholera-related enterotoxin isolated from an Escherichia coli strain of human origin. The immuno-dot-blot assay could detect 0.02 ng of purified enterotoxin. The amount of toxin antigen detected varied widely from strain to strain. Fourteen of the 25 polymyxin B-treated extracts of bacteria harvested from 6-h Casamino Acids-yeast extract broth cultures gave positive results in both serologic assays as well as in rabbit skin tests for delayed permeability factor. An additional strain was positive only in the immuno-dot-blot. Five of six stool isolates and six of seven blood isolates tested gave positive reactions. Two isolates of Salmonella enteritidis tested were also positive. The immuno-dot-blot test appears to be a simple, rapid, and reliable method for detection of cholera-related enterotoxin antigen in S. typhimurium. The demonstration of a cholera-related enterotoxin, even in small amounts, in a facultative intracellular pathogen raises interesting questions regarding its potential role in pathogenesis both of diarrheal disease and systemic infections caused by salmonellae.

摘要

采用先前描述的乳胶颗粒凝集试验以及一种新开发的免疫斑点印迹试验,使用针对从一株人源大肠杆菌分离的霍乱相关肠毒素的免疫纯化山羊抗体,对在印度分离出的25株鼠伤寒沙门氏菌进行霍乱/大肠杆菌相关肠毒素抗原检测。免疫斑点印迹试验能够检测出0.02 ng的纯化肠毒素。不同菌株检测到的毒素抗原量差异很大。从6小时的酪蛋白氨基酸-酵母提取物肉汤培养物中收获的25份经多粘菌素B处理的细菌提取物中,有14份在两种血清学检测以及兔皮试验中对迟发型通透因子呈阳性结果。另外有一个菌株仅在免疫斑点印迹试验中呈阳性。所检测的6份粪便分离株中的5份以及7份血液分离株中的6份呈阳性反应。所检测的2份肠炎沙门氏菌分离株也呈阳性。免疫斑点印迹试验似乎是一种检测鼠伤寒沙门氏菌中霍乱相关肠毒素抗原的简单、快速且可靠的方法。在一种兼性细胞内病原体中即使检测到少量霍乱相关肠毒素,也引发了关于其在沙门氏菌引起的腹泻病和全身感染发病机制中潜在作用的有趣问题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9011/266063/1390cfff99be/jcm00088-0135-a.jpg

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