Department of Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine and College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
College of Animal Science and Technology, Guangdong Polytechnic of Science and Trade, Guangzhou 510640, China.
Poult Sci. 2019 Dec 1;98(12):6492-6496. doi: 10.3382/ps/pez547.
We developed a rapid fluorescent microsphere immunochromatographic test strip (FM-ICTS) assay for the quantitative detection of avian leukosis virus (ALV). A monoclonal antibody specific for the ALV major capsid protein encoded by the gag gene was coupled to label fluorescent microspheres. ALV antibodies were coated on a nitrocellulose membrane to prepare a test line for sample detection. The fluorescence signals of the test and control lines can be read either visually by exposure to UV light or using a fluorescence analyzer. ALV could be detected quantitatively using the ratio of fluorescence signals of the test and control lines (T/C). The assay threshold was determined as a T/C value of 0.0606. The fitting curve equation was established between 1 and 2,048 ng/mL P27 protein with an r2 value of 0.9998. The assay showed no cross reactivity with Newcastle disease virus, infectious laryngotracheitis virus, infectious bronchitis virus, Marek's disease virus, infectious bursal disease, Reoviridae virus, or avian influenza virus. The repeatability was satisfactory with an overall average CV of 8.65%. The Kappa coefficient between a commercial ELISA kit was 0.7031 using clinical chicken meconium samples. Thus, a simple, rapid, sensitive, and specific fluorescent microsphere immunochromatographic test strip was developed based on specific anti-capsid protein p27 monoclonal antibodies.
我们开发了一种快速荧光微球免疫层析检测试剂盒（FM-ICTS），用于定量检测禽白血病病毒（ALV）。一种针对 gag 基因编码的 ALV 主要衣壳蛋白的单克隆抗体被偶联到荧光微球上。ALV 抗体被涂覆到硝酸纤维素膜上，以制备用于样品检测的测试线。可以通过暴露于紫外光下或使用荧光分析仪来读取测试线和控制线的荧光信号。可以通过测试线和控制线的荧光信号比值（T/C）来定量检测 ALV。将检测阈值确定为 T/C 值为 0.0606。在 1 到 2048 ng/mL P27 蛋白之间建立了拟合曲线方程，r2 值为 0.9998。该检测方法与新城疫病毒、传染性喉气管炎病毒、传染性支气管炎病毒、马立克氏病病毒、传染性法氏囊病、呼肠孤病毒或禽流感病毒没有交叉反应。使用临床鸡胎粪样品，重复性令人满意，总平均 CV 为 8.65%。与商业 ELISA 试剂盒的 Kappa 系数为 0.7031。因此，基于特异性抗衣壳蛋白 p27 单克隆抗体，开发了一种简单、快速、灵敏、特异的荧光微球免疫层析检测试剂盒。
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