农业生物技术学报2024，Vol.32Issue(12)：2926-2934,9.DOI:10.3969/j.issn.1674-7968.2024.12.021
间座壳菌属真菌Diaporthe eres的单克隆抗体制备及检测试剂盒研发
Preparation of Monoclonal Antibodies Against Diaporthe eres Fungus and Development of Detection Kit
摘要
Abstract
Fritillaria thunbergii blongs to the genus Fritillaria in the Liliaceae family.The bulb is the main medicinal part,it has the function of clearing heat and dispersing clumps.Leaf spot is one of the main fungal diseases of F.thunbergii,which caused by Diaporthe eres.At present,plant protection personnel and growers mainly rely on classic symptom observation methods to determine the occurrence of leaf spot disease in F.thunbergii,which cannot achieve rapid and accurate diagnosis in its early stages.The purpose of this study was to prepare monoclonal antibodies(McAbs)against D.eres and the immunological detection kits to provide technical support for the rapid and accurate identification of leaf spot in the field.Initially,the protein extract of the hyphae of D.eres was used to immunize Bal b/c mice(Mus musculus).Subsequently,a McAb against D.eres and its hybridoma cell line PoF7 was prepared using hybridoma technology.Indirect ELISA and Western blot analysis showed that the McAb had high specificity against D.eres,with a detection sensitivity of 11 ng/mL.The titer of McAb was≥2.56×104-fold dilution.The antibody type was IgM.It binded to a protein with a molecular weight of 107 kD.The detection sensitivity of the ELISA test kit based on PoF7 was 44 ng/mL,demonstrating good repeatability and specificity.The kit is easy to operate and can be used for large-scale detection of F.thunbergii leaf spot disease in the field after a simple grinding.关键词
浙贝母/叶斑病/Diaporthe eres/单克隆抗体(单抗)/ELISA检测试剂盒Key words
Fritillaria thunbergii/Leaf spot disease/Diaporthe eres/Monoclonal antibody(McAb)/ELISA detection kit分类
农业科技引用本文复制引用
刘强,杭群莺,胡光锦,温思思,赵伟春..间座壳菌属真菌Diaporthe eres的单克隆抗体制备及检测试剂盒研发[J].农业生物技术学报,2024,32(12):2926-2934,9.基金项目
浙江省基础公益研究计划(LGN21C140004) （LGN21C140004）
浙江省大学生科技创新项目(2022R410A037) （2022R410A037）
国家级大学生创新创业训练计划项目(202310344077) （202310344077）
