基于单克隆抗体的酶联免疫吸附测定法检测环境样本中的肠炎沙门氏菌。

Detection of Salmonella enteritidis in environmental samples by monoclonal antibody-based ELISA.

作者信息

Brigmon R L, Zam S G, Bitton G, Farrah S R

机构信息

Department of Environmental Engineering Sciences, University of Florida, Gainesville 32611.

出版信息

J Immunol Methods. 1992 Jul 31;152(1):135-42. doi: 10.1016/0022-1759(92)90096-c.

DOI:10.1016/0022-1759(92)90096-c

PMID:1640107

Abstract

We have developed a enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (ASCII) for the detection of Salmonella enteritidis in environmental samples. ELISA was used to test for sensitivity and specificity of ASCII. 38 other species of bacteria, including 31 Salmonella species were included in cross-reactivity testing with ELISA. ASCII showed no reactivity with any other species tested. ASCII was found to be an IgG1 specific for S. enteritidis lipopolysaccharide (LPS). The lower limits for S. enteritidis detection was 10(5) cells/ml for pure cultures and in 10% sludge (w/v). Environmental samples (raw wastewater, wastewater effluents, mixed liquor and aerobically digested sludge) were obtained twice from five sites and ELISA tested for the presence of S. enteritidis. ELISA results compared to the American Public Health Association (APHA) method of Salmonella detection were not significantly different (P greater than 0.05). The ELISA took 24 h for completion compared to 96-120 h for the APHA procedure. Results demonstrate the reliability of the ELISA and, more importantly, provides a rapid means of detection of S. enteritidis in environmental samples.

摘要

我们已经开发出一种酶联免疫吸附测定法（ELISA），该方法使用单克隆抗体（ASCII）来检测环境样本中的肠炎沙门氏菌。ELISA用于测试ASCII的敏感性和特异性。38种其他细菌，包括31种沙门氏菌，被纳入ELISA的交叉反应性测试。ASCII与所测试的任何其他物种均无反应性。发现ASCII是一种针对肠炎沙门氏菌脂多糖（LPS）的IgG1。纯培养物和10%污泥（w/v）中肠炎沙门氏菌的检测下限为10(5)个细胞/毫升。从五个地点两次采集环境样本（原废水、废水排放物、混合液和需氧消化污泥），并通过ELISA检测肠炎沙门氏菌的存在。与美国公共卫生协会（APHA）检测沙门氏菌的方法相比，ELISA结果无显著差异（P大于0.05）。ELISA完成检测需要24小时，而APHA程序需要96 - 120小时。结果证明了ELISA的可靠性，更重要的是，提供了一种快速检测环境样本中肠炎沙门氏菌的方法。