Highly sensitive detection of Salmonella typhi using surface aminated polycarbonate membrane enhanced-ELISA
Highly sensitive detection of Salmonella typhi using surface aminated polycarbonate membrane enhanced-ELISA
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DOI：
10.1016/j.bios.2011.09.031
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发表时间：
2012-01-15
影响因子：
12.6
通讯作者：
Singh, Harpal
中科院分区：
文献类型：
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作者：
Jain, Swati;Chattopadhyay, Sruti;Singh, Harpal
水中致病菌的鉴定对于解决健康和安全方面的预防和治疗问题非常重要。开发了一种高度灵敏和特异的固相夹心 ELISA 程序，用于在改性 isopore 聚碳酸酯 (PC) 黑膜上检测伤寒引起的极其致命的水传播病原体伤寒沙门氏菌 (S.typhi)。 PC 膜经过化学衍生，在表面生成氨基，保持其物理光学特性。通过ATR-FTIR光谱仪、测角仪和扫描电子显微镜对表面改性PC膜进行了表征。通过胺戊二醛化学，将针对全细胞伤寒沙门氏菌的多克隆体细胞“O”型抗体（Abs）固定在其上。抗体固定膜从缓冲溶液中捕获伤寒沙门氏菌，并使用 HRP 标记的伤寒沙门氏菌抗体通过比色法检测该复合物。使用改性 PC 膜无需任何预富集步骤即可实现 2 x 10(3) 细胞/ml 细菌的检测限，而通过典型 ELISA 方法可实现 10(6)-10(7) CFU/ml 细菌的检测限。与其他交叉反应水源病原体相比，该测定被证明对目标细菌具有特异性。 10(4) 和 10(5) 细胞/ml 的测定内和测定间精度分别为 5.3-7.4 和 10.3-19.7%。与商业上现有的免疫层析测定相比，所开发的用于检测伤寒沙门氏菌的免疫测定法简单、易于操作、灵敏、特异性、可重复性和成本效益。 (C) 2011 Elsevier B.V. 保留所有权利。
The identification of pathogenic bacteria in water is important for addressing preventive and treatment issues regarding health and safety. A highly sensitive and specific solid-phase sandwich ELISA procedure was developed for the detection of typhoid causing extremely lethal water borne pathogen Salmonella typhi (S. typhi) on modified isopore polycarbonate (PC) black membranes. PC membranes were chemically derivatized to generate amino groups on the surface maintaining their pysico-optico properties. Surface modified PC membranes were characterized by ATR-FTIR spectrometer, goniometer and scanning electron microscope. Polyclonal somatic 'O' type antibodies (Abs) against whole cell S. typhi were immobilized on them by following the amine glutaraldehyde chemistry. Antibody immobilized membranes captured S. typhi from buffer solution and this complex was detected colourimetrically using HRP labelled S. typhi Ab. A detection limit of 2 x 10(3) cells/ml of bacteria was achieved with the modified PC membranes without any pre-enrichment step as against 10(6)-10(7) CFU/ml of bacteria by typical ELISA method. The assay was demonstrated to be specific for the target bacteria when compared with other cross-reactant water borne pathogens. The intra- and inter-assay precision for 10(4) and 10(5) cells/ml was 5.3-7.4 and 10.3-19.7% respectively. The developed immunoassay for the detection of S. typhi is simple, easy to handle, sensitive specific, reproducible and cost effective in comparison with the commercially existing immunochromatographic assays. (C) 2011 Elsevier B.V. All rights reserved.