一种新型电化学免疫传感器检测血清4型禽腺病毒  

A novel electrochemical immunosensor for detection of fowl adenovirus serotype 4

作　　者：黄娇玲 谢芝勋 罗思思 李孟 谢丽基 邓显文 谢志勤 曾婷婷 张艳芳 范晴 张民秀 王盛 李丹 韦悠 李小凤 HUANG Jiaoling;XIE Zhixun;LUO Sisi;LI Meng;XIE Liji;DENG Xianwen;XIE Zhiqin;ZENG Tingting;ZHANG Yanfang;FAN Qing;ZHANG Minxiu;WANG Sheng;LI Dan;WEI You;LI Xiaofeng(Key Laboratory of China(Guangxi)-ASEAN Cross-border Animal Disease Prevention and Control of Ministry of Agriculture and Rural Affairs of China/Guangci Key Laboratory of Veterinary Biotechnology,Guangzi Veterinary Research Institute,Nanning 530001,China)

机构地区：[1]广西壮族自治区兽医研究所农业农村部中国(广西)-东盟跨境动物疫病防控重点实验室/广西兽医生物技术重点实验室,广西南宁530001

出　　处：《中国兽医学报》2024年第2期249-257,共9页Chinese Journal of Veterinary Science

基　　金：广西青年科学基金资助项目(2020GXNSFBA297104);广西八桂学者专项基金资助项目(2019A50)。

摘　　要：将多壁碳纳米管-壳聚糖-铂纳米粒子复合材料(MWCNT-Chi-PtNP)修饰金电极(GE),并吸附血清4型禽腺病毒(fowl adenovirus serotype 4,FAdV-4)蛋白单克隆抗体(MAb/FAdV-4),构建一种新型的电阻型电化学免疫传感器用于检测FAdV-4。对传感器的测试底液及样品孵育时间进行优化后,在最优条件下,对所构建的传感器的敏感性、特异性和稳定性进行检测。并对36份临床样品进行检测,以验证该传感器的实用性。结果显示,所建立的FAdV-4电化学免疫传感器的测试底液最佳pH值为7.0,样品的最佳孵育时间为30 min,检测线性范围10^(1.46)~10^(4.46)EID50/mL FAdV-4,最低检出限为10^(1.89)EID50/mL FAdV-4(S/N=3);敏感性试验结果显示该免疫传感器灵敏度比常规PCR方法(10^(3.46)EID50/mL)FAdV-4高37倍,且该传感器特异性高,稳定性好;该传感器对36份临床样品进行检测,结果与PCR方法和测序结果相符。结果表明,所建立的FAdV-4电化学免疫传感器具有检测快速、特异性好和敏感性高等优点,为FAdV-4的诊断提供一种新型的快速检测技术。The aim of this study is to construct a novel resistive electrochemical inmunosensor for the detection of fowl adenovirus serotype 4(FAdV-4).Multi-walled carbon nanotubes-chitosan-Pt nanoparticles(MWCNT-Chi-PtNP)was used to modify gold electrode(GE),then adsorbed with FAdV-4 penton protein monoclonal antibody(MAb/FAdV-4)to construct electrochemical immunosensor for detection of FAdV-4.After optimizing the test substrate and incubation time of the sample,the sensitivity,specificity,and stability of the constructed electrochemical immunosensor were determined under the optimal conditions.Thirty-six clinical samples were detected by the constructed electrochemical immunosensor to verify the practicability of the sensor.Results showed that the optimal pH value of the electrolyte was 7.0,the optimal incubation time of the samples was 30 min.The developed electrochemical immunosensor showed a wide linear range from 10^(1.46)EID_(50)/mL to 10^(4.46)EID_(50)/mL and a detection limit of 10^(1.89)EID_(50)/mL FAdV-4(S/N=3),the sensitivity of the developed electrochemical immunosensor was 37 times higher than that of the conventional PCR(10~(3.46)EID_(50)/mL).This sandwich electrochemical immunosensor also exhibited high selectivity and stability.The results of 36 clinical sample detected by the developed electrochemical immunosensor were consistent with the results of PCR and sequencing.The established electrochemical immunosensor has the advantages of high sensitivity,good specificity,and rapid detection,which provides a new rapid detection for the diagnosis FAdV-4.

关 键 词：多壁碳纳米管 电化学免疫传感器 铂纳米粒子 血清4型禽腺病毒 

分 类 号：S852.6[农业科学—基础兽医学]