中国动物检疫2025，Vol.42Issue(6)：99-105,127,8.DOI:10.3969/j.issn.1005-944X.2025.06.016
鸡传染性喉气管炎病毒抗原双抗夹心ELISA方法的建立
Establishment of a Double-antibody Sandwich ELISA for Avian Infectious Laryngotracheitis Virus
摘要
Abstract
In order to detect avian infectious laryngotracheitis virus(ILTV)pathogen,a double-antibody sandwich ELISA was established using two monoclonal antibodies(mAbs)against ILTV gJ protein,specifically,one mAb served as the capture antibody,while the other was labeled with horseradish peroxidase(HRP)as the detection antibody,and reaction conditions were optimized via checkerboard titration,followed by evaluation on its performance.The results showed that the optimal coating concentration of the capture antibody was 2 μg/mL,and the optimal dilution of the enzyme-labeled mAb was 1:4 000;the critical value was determined to be 0.295 6,the samples pending for test were considered to be positive when the OD450 nm exceeded 0.295 6 and the P/N ratio was larger than 2;the established method could be used to specifically detect ILTV,other than other common avian pathogens;its detection limit was 49.4 ELD50/0.1 mL,and the coefficients of variation of the intra-and-inter assay were less than 10%.For 48 clinical samples,the result of the established method was 93.75%coincident with that of PCR specified by industry standard.In conclusion,the double-antibody sandwich ELISA established in the study could be used to detect ILTV antigen with its advantages of strong specificity,high sensitivity and good repeatability,supporting future detection,monitoring,prevention and control of the disease.关键词
鸡传染性喉气管炎病毒(ILTV)/单克隆抗体/双抗夹心ELISAKey words
ILTV/monocolonal antibody/double-antibody sandwich ELISA分类
农业科技引用本文复制引用
谢晶,毛从剑,杨竣雁,叶健强,曾子轩,康玮笠,康润敏,于吉锋,肖璐,吴学婧,叶勇刚,魏勇,李兴玉,曹冶,潘梦..鸡传染性喉气管炎病毒抗原双抗夹心ELISA方法的建立[J].中国动物检疫,2025,42(6):99-105,127,8.基金项目
四川省区域创新合作计划项目(2024YFHZ0100,2023YFQ0037) （2024YFHZ0100,2023YFQ0037）
四川省财政运行专项(SASA20224CZYX006) （SASA20224CZYX006）
