开发一种灵敏且特异的 xMAP 检测方法，用于检测传染性喉气管炎和传染性支气管炎病毒的抗体。
Development of a sensitive and specific xMAP assay for detection of antibodies against infectious laryngotracheitis and bronchitis viruses.
机构信息
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510640, China.
出版信息
Virol J. 2018 Sep 21;15(1):146. doi: 10.1186/s12985-018-1048-x.
BACKGROUND
A serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination.
METHOD
The microspheres coated with purified recombinant glycoprotein D (gD) of ILTV or nucleocapsid (N) protein of IBV were incubated with serum samples. The simultaneous quantification of ILTV and IBV antibodies were achieved through the interrogation of microspheres by Luminex 200 detection system. .
RESULTS
This xMAP detection demonstrated no nonspecific reactions with avian influenza virus (AIV), avian leukosis virus (ALV), newcastle disease virus (NDV), and Marek's disease virus (MDV). The results also demonstrated that the xMAP assay was four times more sensitive than the enzyme-linked immunosorbent assay (ELISA) for ILTV detection and two times more sensitive for IBV detection. A total of 90 chicken serum samples from a chicken farm were tested by xMAP and ELISA assays. The results showed that the coincidence rates were 84.44 and 100% for ILTV and IBV detection, respectively.
CONCLUSION
This study exhibited an opportunity for the differential diagnosis through simultaneous detection of multiplex antibodies in serum and can be used for the multiplex antibodies evaluation after vaccination.
背景
一种能够同时检测传染性喉气管炎病毒（ILTV）和传染性支气管炎病毒（IBV）抗体的血清学方法对于鉴别诊断和评估疫苗接种后的抗体效价至关重要。
方法
用纯化的 ILTV 糖蛋白 D（gD）或 IBV 核衣壳（N）蛋白包被微球，与血清样品孵育。通过 Luminex 200 检测系统对微球进行检测，实现同时定量检测 ILTV 和 IBV 抗体。
结果
该 xMAP 检测与禽流感病毒（AIV）、禽白血病病毒（ALV）、新城疫病毒（NDV）和马立克氏病病毒（MDV）均无非特异性反应。结果还表明，该 xMAP 检测法对 ILTV 的检测灵敏度比酶联免疫吸附试验（ELISA）高四倍，对 IBV 的检测灵敏度高两倍。用 xMAP 和 ELISA 检测法对来自一个鸡场的 90 份鸡血清样本进行了检测。结果表明，ILTV 和 IBV 的检测符合率分别为 84.44%和 100%。
结论
本研究通过同时检测血清中的多重抗体，为鉴别诊断提供了机会，可用于评估疫苗接种后的多重抗体。
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