Abdelmagid O Y, Mansour M M, Minocha H C, van Drunen Littel-van den Hurk S
Department of Pathobiology, School of Veterinary Medicine, Tuskegee University, AL 36088, USA.
Vet Microbiol. 1998 Apr 15;61(4):249-59. doi: 10.1016/s0378-1135(98)00188-6.
Baculovirus (Autographa californica nuclear polyhedrosis)-expressed bovine herpesvirus-1 (BHV-1) glycoproteins B (gB), gC, and gD were developed and characterized. The recombinant proteins retained their antigenic properties as determined by immunoblotting against monoclonal antibodies. The proteins were examined as antigens for detection of BHV-1 infection and for the analysis of antibody responses to the individual viral proteins. A total of 115 bovine serum samples were tested for their reactivity with individual recombinant proteins from baculovirus-infected Spodoptera frugiperda (SF-9) cell lysates by enzyme-linked immunosorbent assay (ELISA), western blotting, and dot blotting assays. These serum samples were previously tested for BHV-1-specific antibodies by virus neutralization (VN) at the veterinary diagnostic laboratory. All 90 serum samples tested positive with VN were positive by ELISA against gC and gD, separately. However, reactivities of sera against gB were generally low and inconsistent. On the other hand, out of 25 sera that were negative with VN, 22 sera were consistent and gave negative results against gC or gD by ELISA, whereas reactivities with gB were inconsistent. Similar results were obtained when the sera were tested by western blotting and dot blotting. The positive sera consistently reacted strongly against gC and to a lesser extent gD. These results suggest that baculovirus expressed gC from infected cell lysate can be used as a potential diagnostic antigen for detection of anti-gC-specific antibody responses in BHV-1 infected cattle.
开发并鉴定了杆状病毒（苜蓿银纹夜蛾核型多角体病毒）表达的牛疱疹病毒1型（BHV-1）糖蛋白B（gB）、gC和gD。通过针对单克隆抗体的免疫印迹法确定，重组蛋白保留了其抗原特性。对这些蛋白作为抗原用于检测BHV-1感染以及分析针对单个病毒蛋白的抗体反应进行了检测。通过酶联免疫吸附测定（ELISA）、蛋白质印迹法和斑点印迹法，对总共115份牛血清样本与杆状病毒感染的草地贪夜蛾（SF-9）细胞裂解物中的单个重组蛋白的反应性进行了检测。这些血清样本先前在兽医诊断实验室通过病毒中和试验（VN）检测了BHV-1特异性抗体。所有90份VN检测呈阳性的血清样本，通过ELISA分别针对gC和gD检测时也呈阳性。然而，血清对gB的反应性通常较低且不一致。另一方面，在25份VN检测呈阴性的血清中，22份血清结果一致，通过ELISA针对gC或gD检测呈阴性，而与gB的反应性不一致。当通过蛋白质印迹法和斑点印迹法检测血清时，也获得了类似结果。阳性血清始终对gC有强烈反应，对gD的反应程度较小。这些结果表明，杆状病毒从感染细胞裂解物中表达的gC可作为一种潜在的诊断抗原，用于检测BHV-1感染牛中抗gC特异性抗体反应。
