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改性 PAN 纤维免疫分析检测食源性病原体沙门氏菌

Sensitive detection of food-borne pathogen Salmonella by modified PAN fibers-immunoassay.

机构信息

Center for Biomedical Engineering, Indian Institute of Technology, Delhi, India.

出版信息

Biosens Bioelectron. 2013 Jul 15;45:274-80. doi: 10.1016/j.bios.2013.01.032. Epub 2013 Feb 11.

DOI:10.1016/j.bios.2013.01.032
PMID:23500375
Abstract

Sensitive and rapid detection of Salmonella is a key to the prevention and identification of problems associated with human health and safety. Enzyme Linked Immunosorbent Assays (ELISAs) are popular and widely implemented technique to detect pathogenic bacteria in routine analysis but a typical ELISA yields a sensitivity of 10(6)-10(7)cfu/mL. The present study consecrates on the applicability of surface modified polyacrylonitrile (PAN) fibers as a novel matrix of immunoassay for the detection of Salmonella typhimurium in a sandwich ELISA format. Affinity purified antibody against Salmonella common structural antigen (CSA-1-Ab) was immobilized on modified PAN (mPAN) fibers using covalent immobilization via amine-glutaraldehyde chemistry and inactivated S. typhimurium were captured from various samples and detected colorimetrically using peroxidase-labelled common structural antibody (CSA-1-Ab-HRP) against Salmonella. The performance of the developed immunoassay was compared with commercially available immunomagnetic microbeads (Dynabeads(®) anti-Salmonella), polystyrene (PS) microtitre plate and glutaraldehyde activated PS plate. Limit of detection (LOD) was found to be 10, 10(5), 10(6) and 10(7)cells/mL of bacteria for mPAN, Dynabeads(®), glu-plate and PS plate respectively without any pre-enrichment step. The assay was specific for the targeted bacteria when investigated with other cross-reactant food and water-borne pathogens. The developed immunoassay offered undisputed advantages of being simple, sensitive and specific for the detection of S. typhimurium.

摘要

灵敏、快速地检测沙门氏菌是预防和识别与人类健康和安全相关问题的关键。酶联免疫吸附测定(ELISA)是一种广泛应用于常规分析中检测致病菌的流行技术,但典型的 ELISA 灵敏度为 10(6)-10(7)cfu/mL。本研究关注的是表面改性聚丙烯腈(PAN)纤维作为一种新型免疫分析基质在夹心 ELISA 格式中检测鼠伤寒沙门氏菌的适用性。针对沙门氏菌共同结构抗原(CSA-1-Ab)的亲和纯化抗体通过胺-戊二醛化学通过共价固定化固定在改性 PAN(mPAN)纤维上,从各种样品中捕获失活的鼠伤寒沙门氏菌,并使用针对沙门氏菌的过氧化物酶标记共同结构抗体(CSA-1-Ab-HRP)进行比色检测。所开发的免疫测定法的性能与市售免疫磁珠(Dynabeads®抗沙门氏菌)、聚苯乙烯(PS)微量滴定板和戊二醛活化 PS 板进行了比较。发现未经预富集步骤,mPAN、Dynabeads®、glu 板和 PS 板的检测限(LOD)分别为 10、10(5)、10(6)和 10(7)个细菌细胞/mL。当用其他交叉反应的食源性病原体和水源性病原体进行研究时,该测定法对靶细菌具有特异性。所开发的免疫测定法具有简单、灵敏和特异性检测鼠伤寒沙门氏菌的优势。

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