基于免疫磁分离的荧光微球免疫层析法检测猪霍乱沙门氏菌

引用本文: 黄震, 夏诗琪, 刘道峰, 刘成伟, 赖卫华. 基于免疫磁分离的荧光微球免疫层析法检测猪霍乱沙门氏菌[J]. 分析化学, 2017, 45(2): 217-223. doi: 10.11895/j.issn.0253-3820.160572 shu

Citation: HUANG Zhen, XIA Shi-Qi, LIU Dao-Feng, LIU Cheng-Wei, LAI Wei-Hua. Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S. choleraesuis[J]. Chinese Journal of Analytical Chemistry, 2017, 45(2): 217-223. doi: 10.11895/j.issn.0253-3820.160572 shu

基于免疫磁分离的荧光微球免疫层析法检测猪霍乱沙门氏菌

1.
南昌大学食品科学与技术国家重点实验室, 南昌 330047;
2.
江西省林业科学院, 南昌 330032;
3.
江西省疾病预防控制中心, 南昌 330096
基金项目:
本文系国家自然科学基金（No.31271863），江西省教育厅落地项目（No.KJLD13009），江西省生猪产业质量安全岗位专家项目（No.JXARS-03）资助

摘要: 建立了基于免疫磁分离的荧光微球免疫层析法，检测猪霍乱沙门氏菌。待检样品经免疫磁分离富集和热洗脱处理后，用荧光微球免疫层析试纸条进行检测。每毫克纳米磁珠标记30 μg抗体制备的免疫磁珠，对浓度为10²~10⁶ CFU/mL的猪霍乱沙门氏菌的捕获率均大于90%，特异性好；在pH=6时，以300 μg/mg猪霍乱沙门氏菌单抗11D8-D4标记荧光微球，制备免疫荧光微球；以2.0 mg/mL猪霍乱沙门氏菌单抗5F11-B11喷涂检测线（T线），以1.0 mg/mL驴抗鼠IgG喷涂质控线（C线），制备免疫层析试纸条。采用建立的基于免疫磁分离的荧光微球免疫层析方法检测猪霍乱沙门氏菌，在PBS缓冲液中检出限为1.5×10⁵ CFU/mL，牛奶中检出限为7.6×10⁵ CFU/mL，与直接采用荧光微球免疫层析方法检测相比，检出限分别降低了10倍和200倍。本方法可有效富集牛奶中的沙门氏菌，避免了基质干扰，灵敏度大大提高，具有较好的应用前景。

关键词:

English

Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S. choleraesuis

1.
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;
2.
Jiangxi Academy of Forestry, Nanchang 330032, China;
3.
Jiangxi Province Centers for Disease Control and Prevention, Nanchang 330096, China
Received Date: 28 July 2016
Revised Date: 07 December 2016
Fund Project:
This work was supported by the Jiangxi Agriculture Reseach System (No. JXARS-03) and the JIangxi Education Bureau Technology (No. KJLD13009)

Abstract: Immunomagnetic separation (IMS) was coupled with fluorescent microspheres lateral flow assay (FM-LFA) for rapid detection of S. choleraesuis in this study. The target bacteria were firstly enriched from sample by immunomagnetic beads (IMBs), then eluted by heat treatment and detected by fluorescent microspheres lateral flow test strip. The IMBs was labeled with 30 μg/mg antibody, and the capture efficiency was greater than 90% against 10²-10⁶ CFU/mL of S. choleraesuis with great specificity. The immunofluorescent microspheres were prepared by coupling 300 μg of 11D8-D4 monoclonal antibody with 1 mg of fluorescent microspheres at pH 6. Monoclonal antibody 5F11-B11 (2.0 mg/mL) and donkey anti-mouse IgG (1.0 mg/mL) were sprayed on nitrocellulose membrane as test line and control line, respectively. The FM-LFA based on IMS was used to detect S. choleraesuis in PBS and milk. The limits of detection in PBS buffer and milk were 1.5×10⁵ CFU/mL and 7.6×10⁵ CFU/mL respectively, which were 10 and 200 times lower than that of traditional fluorescent microspheres lateral flow assay, respectively. The results showed that the method, which could enrich S. choleraesuis in milk effectively, could avoid matrix interference and improve the detection sensitivity, thus had a good application prospect.

Key words:

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

基于免疫磁分离的荧光微球免疫层析法检测猪霍乱沙门氏菌

English

Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S. choleraesuis

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于免疫磁分离的荧光微球免疫层析法检测猪霍乱沙门氏菌

English

Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S. choleraesuis

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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