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鸡和鸭新城疫病毒抗体竞争酶联免疫吸附试验的研制及田间应用。

Development and field application of a competitive enzyme-linked immunosorbent assay for detection of Newcastle disease virus antibodies in chickens and ducks.

机构信息

Department of Microbiology and Infectious Disease, Hanoi University of Agriculture, Hanoi, Vietnam.

出版信息

Poult Sci. 2013 Aug;92(8):2034-43. doi: 10.3382/ps.2013-03176.

DOI:10.3382/ps.2013-03176
PMID:23873550
Abstract

A competitive enzyme-linked immunosorbent assay (C-ELISA) using a baculovirus-expressed recombinant nucleocapsid protein antigen (rNDV-N) and an rNDV-N-specific monoclonal antibody (5B3) was developed for the detection of Newcastle disease virus (NDV) antibodies, and its diagnostic performance was evaluated. The specificity and sensitivity of the C-ELISA was found to be 98.4 and 98.9%, respectively, for chickens, and 98.2 and 97.9% for ducks. However, the C-ELISA showed weak cross-reaction with hyperimmune antisera to some other avian paramyxovirus serotypes. In all experimentally vaccinated chickens, seroconversion rates at 7 d postinoculation were 100 and 40% when measured by C-ELISA and hemagglutination inhibition (HI), respectively. In field trials, the C-ELISA showed positive results in 98.9% of HI-positive sera and 40.8% of HI-negative sera from NDV-vaccinated chickens (n = 705). In domestic ducks (n = 158) from NDV-positive duck farms (n = 8), the positive rates according to C-ELISA were significantly higher than those according to the HI test. At the same time, 98.1% of ducks (n = 209) from NDV-negative duck farms (n = 11) were also negative by C-ELISA. Our results indicate that C-ELISA could be a useful alternative to HI testing for detecting NDV antibodies in different avian species such as chickens and ducks.

摘要

建立了一种基于杆状病毒表达的重组核衣壳蛋白抗原(rNDV-N)和 rNDV-N 特异性单克隆抗体(5B3)的竞争酶联免疫吸附试验(C-ELISA),用于检测新城疫病毒(NDV)抗体,并评估了其诊断性能。该 C-ELISA 对鸡的特异性和敏感性分别为 98.4%和 98.9%,对鸭的特异性和敏感性分别为 98.2%和 97.9%。然而,该 C-ELISA 与某些其他禽副黏病毒血清型的高免血清显示出较弱的交叉反应。在所有经实验接种的鸡中,C-ELISA 和血凝抑制(HI)分别在接种后 7 天的血清转化率为 100%和 40%。在田间试验中,C-ELISA 在 HI 阳性血清中的阳性率为 98.9%,在 HI 阴性血清中的阳性率为 40.8%(n=705)。在来自 NDV 阳性鸭场的 158 只家鸭(n=8)中,根据 C-ELISA 的阳性率明显高于根据 HI 试验的阳性率。同时,来自 NDV 阴性鸭场的 98.1%的鸭子(n=209)(n=11)也为阴性。我们的结果表明,C-ELISA 可以替代 HI 试验,用于检测不同禽类如鸡和鸭中的 NDV 抗体。

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