农业生物技术学报2025，Vol.33Issue(1)：220-227,8.DOI:10.3969/j.issn.1674-7968.2025.01.020
基于核苷三磷酸二磷酸水解酶的马锥虫病间接ELISA抗体检测方法的建立
Establishment of an Indirect ELISA Antibody Detection Method for Equine Trypanosomosis Based on Nucleotide Triphosphate Diphosphohydrolase
摘要
Abstract
Trypanosomosis(Surra)by Trypanosoma evansi,which resides in the blood,is a protozoan disease in animals such as horses(Equus)and camels(Camelus).Accurate diagnosis is a crucial step in the effective prevention and control of Surra,but the existing diagnostic techniques have many drawbacks such as high false negative rates and high prices.Based on recombinant expression of nucleoside triphosphate diphosphate hydrolase(TeNTPDase)from T.evansi,in this study,an ELISA method was established for Surra by optimizing various reaction parameters,including antigen coating concentration and conditions,serum dilution and incubation conditions,conjugated second antibody dilution and incubation conditions,and evaluated its diagnostic efficacy.The results showed that the recombinant TeNTPDase was mainly expressed in the form of inclusion bodies,with a size of approximately 60 kD.The coefficient of variation of the established ELISA method in different batches of coating antigens were 3.50％～7.32％being less than 10％,and the coefficient of variation at different incubation time points at 37℃ were 3.89％～14.45％being less than 15％,indicating good repeatability and stability.Moreover,when using the established ELISA method to detect clinical serum samples of horses,it was found that the positive rate of Surra was 5.83％(7/120),slightly higher than that of the commercial test kit(3.33％),but the difference was not significant,with a coincidence rate of 92.5％(111/120).This study provides technical support for the effective prevention and control of Surra in horses.关键词
伊氏锥虫病/马/核苷三磷酸二磷酸水解酶(NTPDase)/ELISAKey words
Surra/Horse/Nucleoside triphosphate diphosphohydrolase(NTPDase)/ELISA分类
农业科技引用本文复制引用
曾晴勤,郑亚东,赵秀玲,宋厚辉,李如松,李红英,王欣乐,王莹,夏天奇,王璞,时恒枝,张路平..基于核苷三磷酸二磷酸水解酶的马锥虫病间接ELISA抗体检测方法的建立[J].农业生物技术学报,2025,33(1):220-227,8.基金项目
海关总署科研计划项目(2022HK040) （2022HK040）
宁波中盛产品检测有限公司技术开发项目(2023ZS003) （2023ZS003）
上海市加强公共卫生体系建设三年行动计划(2023-2025年)重点学科项目(GWVI-11.1-12) （2023-2025年）
浙江农林大学科研发展基金项目(2022LFR035) （2022LFR035）
