双重放大酶免疫传感器检测鸡白痢鸡伤寒沙门菌

畜牧兽医学报

双重放大酶免疫传感器检测鸡白痢鸡伤寒沙门菌

胡春梅，窦文超，赵广英^*

（浙江工商大学食品与生物工程学院浙江省食品安全重点实验室，杭州310035）

收稿日期:2012-06-18 出版日期:2012-12-26 发布日期:2012-12-22
通讯作者: 赵广英，教授，E-mail:zhaoguangying18@163.com
作者简介:胡春梅(1987-)，女，四川南充人，硕士生，主要从事食品质量与安全研究，E-mail:dandelionhcm@163.com
基金资助:
国家自然科学基金资助项目(30571623);浙江工商大学校级创新基金(1110XJ1511104)；浙江省大学生科技推广项目(1110JQ4212048G)

Enzyme Immunosensor of Double Amplification for Detection of Salmonella pullorum and Salmonella gallinarum

HU Chun-mei, DOU Wen-chao, ZHAO Guang-ying^*

(Food Safety Key Laboratory of Zhejiang Province, College of Food Science and Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou 310035, China)

Received:2012-06-18 Online:2012-12-26 Published:2012-12-22

摘要/Abstract

摘要：

为探求新型鸡白痢鸡伤寒沙门菌快速检测技术，本研究首次利用多壁碳纳米管与离子液体［BMIM］PF₆修饰四通道丝网印刷碳电极，制成双重放大信号检测鸡白痢鸡伤寒沙门菌的酶免疫传感器。用原子力显微镜表征其表观形态，循环伏安法监测其电化学特性。结果表明，在优化的检测条件下，免疫电极对目标菌的线性响应范围为10³～10⁹ cfu·mL^－1，检出限为3.93×10² cfu·mL^－1（S/N=3），4 ℃放置28 d后，响应电流为初始值的90.15%，具有良好的稳定性、特异性、重现性和准确性。多壁碳纳米管与离子液体结合制备的鸡白痢鸡伤寒沙门菌酶免疫传感器实现了检测信号的双重放大，检测灵敏度高，离子液体有助于保持酶标抗体的活性，延长免疫电极的有效时间。依据该方法构建的酶免疫传感器为快速检测其它致病菌酶免疫传感器的研究提供了良好的参照模型。

关键词: 快速检测, 鸡白痢鸡伤寒沙门菌, 酶免疫传感器, 双重放大, 多壁碳纳米管, 离子液体

Abstract:

The aim of this work is to seek a new rapid detection technology for Salmonella pullorum and Salmonella gallinarum. In this report, we firstly combined multiwalled carbon nanotubes with ionic liquid ［BMIM］PF₆ to modify four-channel screen-printed carbon electrode to propose an enzyme immunosensor with double amplified response signal for detection of Salmonella pullorum and Salmonella gallinarum. The surface morphology of the modified electrode was characterized by atomic force microscopy, while the electrochemical properties were determined by cyclic voltammetry. The results showed that under the optimal assay conditions, a good linear response occurred in the concentration range of 10³-10⁹ cfu·mL^－1 of Salmonella pullorum and Salmonella gallinarum, with a low detection limit of 3.93×10² cfu·mL^－1 (S/N=3). The stability test of the fabricated immunosensor showed that it remained 90.15% of its original response signal after storage of 28 days under 4 ℃. The performance evaluation of the proposed immunosensor showed that it had high stability, good specificity, acceptable reproducibility and accuracy. The results demonstrated that the proposed enzyme immunosensor with combination of multi-walled carbon nanotubes and ionic liquid achieved the effect of double amplification of detection signal which enabled it highly sensitive in detection. Ionic liquid was also helpful for maintaining bioactivity of enzyme labeled antibody, thereby prolonging effective time of the immune electrode. The fabrication of the enzyme immunosensor pursuant to this method provides a good reference model for construction of enzyme immunosensor for rapid detection of other pathogenic bacterium.

Key words: rapid detection, Salmonella pullorum and Salmonella gallinarum, enzyme immunosensor, double amplification, multi-walled carbon nanotubes, ionic liquid

中图分类号:

胡春梅，窦文超，赵广英. 双重放大酶免疫传感器检测鸡白痢鸡伤寒沙门菌[J]. 畜牧兽医学报.

HU Chun-mei, DOU Wen-chao, ZHAO Guang-ying. Enzyme Immunosensor of Double Amplification for Detection of Salmonella pullorum and Salmonella gallinarum[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA.

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