Mahmoodi Pezhman, Seyfi Abad Shapouri Masoud Reza, Ghorbanpour Masoud, Haji Hajikolaei Mohammad Rahim, Lotfi Mohsen, Pourmahdi Boroujeni Mahdi, Daghari Maryam
Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, IR Iran.
Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, IR Iran.
Jundishapur J Microbiol. 2015 Jan 14;8(3):e14311. doi: 10.5812/jjm.14311. eCollection 2015 Mar.
Bovine viral diarrhea (BVD) is an economically important disease of cattle distributed worldwide. Diagnosis of BVD relies on laboratory-based detection of its viral causing agent or virus specific antibodies and the most common laboratory method for this purpose is Enzyme-Linked Immunosorbent Assay (ELISA).
The current study was aimed to develop a simple indirect ELISA to detect antibodies against Bovine Viral Diarrhea Virus (BVDV) in the sera of infected cattle.
A new simple indirect ELISA method was developed to detect BVDV infection by prokaryotically (Escherichia coli, BL21 strain) expressed recombinant whole nonstructural protein 3 (NS3) of BVDV (NADL strain). Four hundred bovine serum samples were evaluated by the newly developed NS3-ELISA and virus neutralization test (VNT) as the gold standard method to diagnose BVD. Among these samples, 289 sera had been previously tested by a commercial ELISA kit.
Statistical analyses showed a very high correlation between the results of the developed NS3-ELISA and VNT (kappa coefficient = 0.935, P < 0.001), with the relative sensitivity and specificity of 94% and 98.8%, respectively. There was also a high correlation between the results of NS3-ELISA and the commercial ELISA kit (kappa coefficient = 0.802, P < 0.001) with the relative sensitivity and specificity of 90.72% and 91.15%, respectively.
The newly developed simple indirect ELISA showed high sensitivity and specificity with respect to VNT. Developing such a simple, sensitive, and specific ELISA which is much less expensive than the available commercial ELISA kits can improve the detection of BVDV infections, help to eliminate the disease from herds, and decrease economic losses caused by this disease.
牛病毒性腹泻（BVD）是一种在全球范围内分布的对牛具有重要经济影响的疾病。BVD的诊断依赖于基于实验室的病毒病原体或病毒特异性抗体检测，为此最常用的实验室方法是酶联免疫吸附测定（ELISA）。
本研究旨在开发一种简单的间接ELISA方法，用于检测感染牛血清中抗牛病毒性腹泻病毒（BVDV）的抗体。
开发了一种新的简单间接ELISA方法，通过原核（大肠杆菌，BL21菌株）表达的BVDV（NADL株）重组全非结构蛋白3（NS3）来检测BVDV感染。采用新开发的NS3-ELISA和作为诊断BVD金标准方法的病毒中和试验（VNT）对400份牛血清样本进行评估。在这些样本中，289份血清此前已用商用ELISA试剂盒检测过。
统计分析表明，所开发的NS3-ELISA与VNT结果之间具有非常高的相关性（kappa系数 = 0.935，P < 0.001），相对敏感性和特异性分别为94%和98.8%。NS3-ELISA与商用ELISA试剂盒结果之间也具有高度相关性（kappa系数 = 0.802，P < 0.001），相对敏感性和特异性分别为90.72%和91.15%。
新开发的简单间接ELISA相对于VNT显示出高敏感性和特异性。开发这样一种比现有商用ELISA试剂盒便宜得多的简单、灵敏且特异的ELISA，可改善BVDV感染的检测，有助于从牛群中消除该疾病，并减少该疾病造成的经济损失。
