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胶体金检测试纸条的研制及其在传染性喉气管炎病毒快速检测中的应用。

Development and application of a colloidal gold test strip for the rapid detection of the infectious laryngotracheitis virus.

机构信息

Sichuan Animal Science Academy, Sichuan Provincial Key Laboratory of Animal Breeding and Genetics, Chengdu 610066, China.

College of Life Science and Technology, Southwest Minzu University, Chengdu, 610041, China.

出版信息

Poult Sci. 2020 May;99(5):2407-2415. doi: 10.1016/j.psj.2019.11.066. Epub 2020 Mar 17.

Abstract

Infectious laryngotracheitis disease is an acute, highly contagious viral disease seriously affecting poultry industry worldwide. In this study, a rapid and simple immune colloidal gold test strip for detecting infectious laryngotracheitis virus (ILTV) was developed based on membrane chromatography with monoclonal antibodies (mAbs) against gJ protein of ILTV and systematically evaluated for the detection of ILTV from clinical samples. mAb 2D4 1D7 was conjugated with colloidal gold as the detector antibody on the test strip. Another mAb, 1D8 1G3, was used as the capture complex at the test line (T-line), and goat antimouse IgG antibody was used as the capture antibody at the control line (C-line). The colloidal gold test strip showed high specificity in the detection of ILTV, with no cross-reaction with other avian pathogens, including infectious bronchitis virus, infectious bursal disease virus, avian influenza virus, Newcastle disease virus, fowl adenoviruses, and Marek's disease virus. Besides, the detection limit of this method was as low as 60 ELD/mL for the ILTV Wanggang strain. Furthermore, we evaluated its application in 260 clinical samples suspected of infection with ILTV. Results from the strip test were nearly identical with those from real-time PCR (coincidence rate 99.6%) and showed higher sensitivity than conventional PCR. All the results obtained in this study indicated that the colloidal gold test strip can be applied as a simple, rapid, sensitive, and specific diagnostic tool for the detection of ILTV, especially in resource-limited areas.

摘要

传染性喉气管炎病是一种急性、高度传染性的病毒性疾病,严重影响着全球的家禽养殖业。本研究基于膜色谱法,利用针对传染性喉气管炎病毒(ILTV)gJ 蛋白的单克隆抗体(mAbs),开发了一种用于检测 ILTV 的快速简便免疫胶体金测试条,并对其从临床样本中检测 ILTV 的能力进行了系统评估。mAb 2D4 1D7 与胶体金缀合作为测试带上的检测抗体。另一种 mAb,1D8 1G3,被用作测试线(T 线)上的捕获复合物,而山羊抗小鼠 IgG 抗体被用作控制线(C 线)上的捕获抗体。胶体金测试条在检测 ILTV 时表现出高度的特异性,与其他禽病病原体(包括传染性支气管炎病毒、传染性法氏囊病病毒、禽流感病毒、新城疫病毒、禽腺病毒和马立克氏病病毒)无交叉反应。此外,该方法的检测限低至 60 ELD/mL 的 ILTV Wanggang 株。此外,我们评估了其在 260 份疑似感染 ILTV 的临床样本中的应用。该条带测试的结果与实时 PCR 几乎一致(符合率为 99.6%),且比传统 PCR 具有更高的灵敏度。本研究的所有结果表明,胶体金测试条可作为一种简单、快速、灵敏、特异的 ILTV 检测诊断工具,尤其适用于资源有限的地区。

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