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用于检测霍乱弧菌O139的5分钟快速检测方法的研发。

Development of a 5-minute rapid test for detecting Vibrio cholerae O139.

作者信息

Thattiyaphong A, Okada K, Khangrang S, Nispa W, Sawanpanyalert P, Honda T

机构信息

National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.

出版信息

Southeast Asian J Trop Med Public Health. 2013 May;44(3):448-55.

Abstract

Vibrio cholerae O139 is an etiology of cholera in Thailand. We determined to prepare a rapid test to detect V cholerae O139 using an immunochromatographic method to be used for surveillance and use in community laboratories. We conjugated murine monoclonal antibodies specific to the lipopolysaccharide of V. cholerae O139 with colloidal gold particles. The sensitivity of the test was determined using 10-fold dilutions of V. cholerae O139. The lowest number of bacterial cells detected by the test was 10(6) cfu/ml. The specificity was determined using 51 isolates of pure cultures, including V. cholerae serogroups O1, O139, and non-O1/non-O139 and other enteric bacteria; 27 rectal swab specimens and 100 specimens of enriched alkaline peptone water, 50 of which contained V. cholerae O1 and the other did not contain V. cholerae O1. The rapid test showed specificity to V cholerae O139 and no cross reaction with V cholerae O1, V. cholerae non-O1/ non-O139, and other enteric bacteria. The test is rapid, simple and easy to use.

摘要

霍乱弧菌O139是泰国霍乱的病因之一。我们决定采用免疫层析法制备一种用于检测霍乱弧菌O139的快速检测方法,以用于监测并在社区实验室使用。我们将针对霍乱弧菌O139脂多糖的鼠单克隆抗体与胶体金颗粒偶联。使用霍乱弧菌O139的10倍稀释液测定该检测方法的灵敏度。该检测方法检测到的最低细菌细胞数为10(6) cfu/ml。使用51株纯培养物测定特异性,包括霍乱弧菌O1、O139血清群、非O1/非O139血清群以及其他肠道细菌;27份直肠拭子标本和100份增菌碱性蛋白胨水标本,其中50份含有霍乱弧菌O1,另外50份不含霍乱弧菌O1。该快速检测方法对霍乱弧菌O139具有特异性,与霍乱弧菌O1、非O1/非O139霍乱弧菌以及其他肠道细菌无交叉反应。该检测方法快速、简单且易于使用。

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