Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Guangdong Laboratory Animals Monitoring Institute, Guangzhou 510640, China.
Poult Sci. 2019 Mar 1;98(3):1494-1499. doi: 10.3382/ps/pey510.
Using Luminex xMAP (x = analyte, MAP = multi-analyte profiling) technology, a serological method for the simultaneous detection of antibodies to Newcastle disease virus (NDV) and avian influenza virus (AIV) was established. Nano-magnetic beads coated with purified NDV protein and AIV nucleoprotein were incubated with serum samples. Using biotinylated rabbit anti-chicken IgY and streptavidin-R-phycoerythrin, the optical signals measured by a Luminex 200 detection system indicated the quantification of NDV or AIV antibodies in the serum. Specific pathogen-free (SPF) chicken serum was used as a negative control. The Luminex xMAP assay developed in this study demonstrated high specificity as there was no cross-reaction with antibodies to infectious laryngotracheitis virus, infectious bronchitis virus, infectious bursal disease virus, avian leukosis virus, and Marek's disease virus. The results from reproducibility experiments showed that intra-coefficients of variation were 3.36 and 9.23% and inter-coefficients of variation were 6.50 and 7.66% for NDV and AIV, respectively. The results also indicated that the Luminex xMAP assay was 16 times more sensitive for NDV antibody detection and 1,024 times more sensitive for AIV antibody detection compared to the enzyme-linked immunosorbent assay (ELISA). A total of 300 chicken serum samples were subjected to both Luminex xMAP assay and ELISA, showing the coincidence rates of 98.67 and 98% for NDV and AIV antibody detection, respectively. This study provides a new method for the simultaneous detection NDV and AIV antibodies in the serum with high specificity and sensitivity.
利用 Luminex xMAP（x = 分析物，MAP = 多分析物分析）技术，建立了一种同时检测新城疫病毒（NDV）和禽流感病毒（AIV）抗体的血清学方法。将纯化的 NDV 蛋白和 AIV 核蛋白包被的纳米磁珠与血清样本孵育。使用生物素化兔抗鸡 IgY 和链霉亲和素-R-藻红蛋白，通过 Luminex 200 检测系统测量的光学信号表明血清中 NDV 或 AIV 抗体的定量。无特定病原体（SPF）鸡血清用作阴性对照。本研究中开发的 Luminex xMAP 测定法表现出很高的特异性，因为与传染性喉气管炎病毒、传染性支气管炎病毒、传染性法氏囊病病毒、禽白血病病毒和马立克氏病病毒的抗体没有交叉反应。重复性实验的结果表明，NDV 和 AIV 的内变异系数分别为 3.36%和 9.23%，而间变异系数分别为 6.50%和 7.66%。结果还表明，与酶联免疫吸附测定（ELISA）相比，Luminex xMAP 测定法检测 NDV 抗体的灵敏度提高了 16 倍，检测 AIV 抗体的灵敏度提高了 1024 倍。对 300 份鸡血清样本同时进行 Luminex xMAP 测定法和 ELISA 检测，NDV 和 AIV 抗体检测的符合率分别为 98.67%和 98%。本研究提供了一种同时检测血清中 NDV 和 AIV 抗体的新方法，具有高特异性和敏感性。
