Establishment of xMAP for the simultaneous detection of antibodies to Newcastle disease virus and avian influenza virus
Establishment of xMAP for the simultaneous detection of antibodies to Newcastle disease virus and avian influenza virus
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新城疫病毒和禽流感病毒抗体同时检测xMAP的建立
DOI：
10.3382/ps/pey510
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发表时间：
2019-03-01
期刊：
影响因子：
4.4
通讯作者：
Guo, Pengju
中科院分区：
文献类型：
--
作者：
Wang, Huanan;Cong, Feng;Guo, Pengju
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。将包被纯化的NDV蛋白和AIV核蛋白的纳米磁珠与血清样本一起孵育。利用生物素化的兔抗鸡IgY和链霉亲和素 - R - 藻红蛋白，通过Luminex 200检测系统测量的光信号表明了血清中NDV或AIV抗体的定量。无特定病原体（SPF）鸡血清用作阴性对照。本研究开发的Luminex xMAP检测方法显示出高特异性，因为它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无交叉反应。重复性实验结果表明，NDV和AIV的组内变异系数分别为3.36%和9.23%，组间变异系数分别为6.50%和7.66%。结果还表明，与酶联免疫吸附测定（ELISA）相比，Luminex xMAP检测方法对NDV抗体检测的灵敏度高16倍，对AIV抗体检测的灵敏度高1024倍。总共300份鸡血清样本同时进行了Luminex xMAP检测和ELISA检测，NDV和AIV抗体检测的符合率分别为98.67%和98%。本研究为同时检测血清中的NDV和AIV抗体提供了一种具有高特异性和灵敏度的新方法。
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.