用于牛疱疹病毒5血清学诊断的间接酶联免疫吸附测定法的开发
Development of an Indirect ELISA for Serological Diagnosis of Bovine herpesvirus 5.
作者信息
Dummer Luana A, Araujo Itauá L, Campos Fabrício S, da Rosa Matheus C, Finger Paula F, de Oliveira Patricia D, Conceição Fabricio R, Fischer Geferson, Roehe Paulo M, Leite Fábio P L
机构信息
Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Laboratório de Virologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
出版信息
PLoS One. 2016 Feb 11;11(2):e0149134. doi: 10.1371/journal.pone.0149134. eCollection 2016.
Bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) are economically important pathogens, associated with a variety of clinical syndromes, including respiratory and genital disease, reproductive failure and meningoencephalitis. The standard serological assay to diagnose BoHV-1 and BoHV-5 infections is the virus neutralization test (VNT), a time consuming procedure that requires manipulation of infectious virus. In the present study a highly sensitive and specific single dilution indirect ELISA was developed using recombinant glycoprotein D from BoHV-5 as antigen (rgD5ELISA). Bovine serum samples (n = 450) were screened by VNT against BoHV-5a and by rgD5ELISA. Compared with the VNT, the rgD5ELISA demonstrated accuracy of 99.8%, with 100% sensitivity, 96.7% specificity and coefficient of agreement between the tests of 0.954. The rgD5ELISA described here shows excellent agreement with the VNT and is shown to be a simple, convenient, specific and highly sensitive virus-free assay for detection of serum antibodies to BoHV-5.
牛疱疹病毒1型和5型（BoHV-1和BoHV-5）是具有重要经济意义的病原体，与多种临床综合征相关，包括呼吸道和生殖道疾病、生殖失败和脑膜脑炎。诊断BoHV-1和BoHV-5感染的标准血清学检测方法是病毒中和试验（VNT），这是一种耗时的程序，需要处理传染性病毒。在本研究中，使用来自BoHV-5的重组糖蛋白D作为抗原开发了一种高度敏感和特异的单稀释间接ELISA（rgD5ELISA）。用VNT检测针对BoHV-5a的牛血清样本（n = 450），并用rgD5ELISA进行检测。与VNT相比，rgD5ELISA的准确率为99.8%，灵敏度为100%，特异性为96.7%，两种检测方法之间的一致性系数为0.954。这里描述的rgD5ELISA与VNT显示出极好的一致性，并且被证明是一种用于检测牛血清中BoHV-5抗体的简单、方便、特异且高度敏感的无病毒检测方法。
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