烟草花叶病毒胶体金检测试纸条的制备与性能评价  

Preparation and performance evaluation of colloidal gold test strips for tobacco mosaic virus detection

作　　者：王春琼 刘凯 谢永辉[2] 张轲 刘春明[3] 张晓伟 孙浩巍 陈丹 WANG Chunqiong;LIU Kai;XIE Yonghui;ZHANG Ke;LIU Chunming;ZHANG Xiaowei;SUN Haowei;CHEN Dan(Yunnan Tobacco Quality Supervision and Test Station,Kunming 650106,China;Yunnan Tobacco Company Kunming City Company,Kunming 650000,China;Yunnan Tobacco Company Honghe Prefecture,Mile 652300,China)

机构地区：[1]云南省烟草质量监督检测站,云南昆明650106 [2]云南省烟草公司昆明市公司,云南昆明650000 [3]云南省烟草公司红河州公司,云南弥勒652300

出　　处：《轻工学报》2025年第1期90-97,共8页Journal of Light Industry

基　　金：云南省烟草专卖局重大科技项目(2021530000241021)。

摘　　要：针对传统烟草花叶病毒(TMV)检测方法耗时较长、应用受限的问题,基于双抗夹心法研发快速定性检测TMV的胶体金检测试纸条,并对其制备条件进行优化,考查其检测限、准确性和特异性。结果表明:该试纸条的适宜制备条件为缓冲液添加量4μL、抗体质量浓度10μg/mL、NC膜选择CN140型、样本提取液0.02 mol/L PBS;该试纸条对TMV-CP蛋白的检测限为0.1μg/mL,与烟草丛顶病毒、番茄斑萎病毒、马铃薯Y病毒无交叉反应,特异性良好;将其进行实际样品检测,与PCR的总符合率为95.0%,阳性符合率为93.3%,准确性良好。此试纸条在10 min内即可显示检测结果,且操作简单,可用于烟田TMV的快速检测。To address the problems of the time-consuming and limited application of traditional tobacco mosaic virus(TMV)detection methods,colloidal gold test strips for rapid and qualitative detection of TMV were developed based on the double antibody sandwich method,and its preparation conditions were optimized.In addition,the limit,accuracy,and specificity of the test strip were detected.The results showed that the suitable preparation conditions for the test strip were buffer addition of 4μL,antibody mass concentration of 10μg/mL,and NC membrane of CN140 type and sample extraction solution of 0.02 mol/L PBS;The detection limit of this test strip for TMV-CP protein was 0.1μg/mL,with no cross reactivity with tobacco bush top virus,tomato spot wilt virus,and potato Y virus,and good specificity;Applied to actual sample detection,the total coincidence rate with PCR was 95.0%,the positive coincidence rate was 93.3%,and the accuracy was good.This test strip could display the detection results within 10 min and was easy to operate.It could be used for rapid detection of TMVs in tobacco field.

关 键 词：烟草花叶病毒 单克隆抗体 胶体金免疫层析技术 试纸条 

分 类 号：S432.41[农业科学—植物病理学]