Indirect ELISA developed to detect antibodies against Mycoplasma synoviae P50 protein via immunoproteomic screening
文献类型： 外文期刊
作者： Liu, Yang 1 ; You, Guangju 1 ; Shi, Jialei 1 ; Gao, Li 1 ; Li, Xiaoqi 1 ; Cao, Hong 1 ; Wang, Yongqiang 1 ; Zheng, Shijun J. 1 ;
作者机构： 1.China Agr Univ, Coll Vet Med, Natl Key Lab Vet Publ Hlth Secur, Beijing, Peoples R China
2.China Agr Univ, Coll Vet Med, Key Lab Anim Epidemiol, Minist Agr, Beijing, Peoples R China
3.China Agr Univ, Coll Vet Med, Beijing, Peoples R China
4.Fujian Acad Agr Sci, Inst Anim Husb & Vet Med, Lab Anim Virol, Fuzhou, Peoples R China
关键词： Mycoplasma synoviae; Lipoprotein; P50 protein; Indirect ELISA
期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.3； 五年影响因子：5.1 ）
ISSN： 0175-7598
年卷期： 2025 年 109 卷 1 期
页码：
收录情况： SCI
摘要： Mycoplasma synoviae infection is a chronic disease of poultry with significant economic impacts. An efficient diagnostic tool for M. synoviae infection is in great demand. This study aimed to develop a novel indirect enzyme-linked immunosorbent assay (iELISA) method based on antigens identified by pull-down assay combined with mass spectrometry. Using these methods and anti-M. synoviae serum, we identified an uncharacterized protein with a molecular weight of 53 kDa (named P50 protein) and then established a recombinant P50 protein-based ELISA (rP50-ELISA) to detect antibodies against P50 protein. A receiver operating characteristic (ROC) analysis was performed to estimate the optical density (OD) cut-off value that maximized the sensitivity (Se) and specificity (Sp) of the rP50-ELISA, which had a mean Se of 93% (95% confidence interval (CI) = 86.25-96.57%) and a mean Sp of 100% (95% CI = 91.80-100%), with an area under the curve (AUC) of 0.9979 (95% CI = 99.41-100%). The rP50-ELISA showed no cross-reactivity with antibodies against other avian pathogens. Serum samples from 164 clinical chickens were tested with the rP50-ELISA, and the results revealed a high concordance rate of 93.29% with commercial diagnostic kits.
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