Letchworth G J, Fishel J R, Hansen W
Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
Avian Dis. 1997 Oct-Dec;41(4):808-16.
Inclusion body disease of cranes (IBDC) herpesvirus kills some infected cranes and persists in convalescent animals. To enable further study and rapid identification of carrier animals, we developed a monoclonal antibody (MAb) to IBDC virus and used it in immunohistochemistry and a competitive enzyme-linked immunosorbent assay (ELISA). We used conventional techniques to make murine MAbs directed against IBDC virus purified from infected duck embryo cells. Hybridomas reacting in an ELISA with IBDC virus but not uninfected duck embryo cells were characterized by radioimmunoprecipitation, in situ immunohistochemistry, and competitive ELISA with neutralizing and nonneutralizing crane sera. MAb 2C11 immunoprecipitated 59-, 61-, and 110-kD proteins from IBDC virus-infected but not uninfected cells and stained glutaraldehyde-fixed IBDC virus plaques but not surrounding uninfected duck embryo cells in vitro. Antibody 2C11 did not react with duck embryo cells infected with falcon herpesvirus, psittacine herpesvirus, infectious laryngotracheitis, pigeon herpesvirus, or duck plague virus. A competitive ELISA using antibody 2C11 identified most sera that were positive in the neutralization test. This antibody will be useful in further characterizing IBDC virus, its pathogenesis, and its natural history.
鹤包涵体病（IBDC）疱疹病毒可导致部分感染鹤死亡，并在康复动物体内持续存在。为了进一步研究并快速鉴定携带病毒的动物，我们制备了一种针对IBDC病毒的单克隆抗体（MAb），并将其用于免疫组织化学和竞争性酶联免疫吸附测定（ELISA）。我们采用常规技术制备了针对从感染鸭胚细胞中纯化得到的IBDC病毒的鼠单克隆抗体。通过放射免疫沉淀、原位免疫组织化学以及使用中和及非中和鹤血清的竞争性ELISA，对在ELISA中与IBDC病毒发生反应但不与未感染鸭胚细胞发生反应的杂交瘤进行了鉴定。单克隆抗体2C11能从感染IBDC病毒的细胞而非未感染细胞中免疫沉淀出59-kD、61-kD和110-kD的蛋白质，并且在体外可对戊二醛固定的IBDC病毒蚀斑进行染色，但不染色周围未感染的鸭胚细胞。抗体2C11不与感染隼疱疹病毒、鹦鹉疱疹病毒、传染性喉气管炎、鸽疱疹病毒或鸭瘟病毒的鸭胚细胞发生反应。使用抗体2C11的竞争性ELISA可鉴定出中和试验中呈阳性的大多数血清。这种抗体将有助于进一步阐明IBDC病毒、其发病机制及其自然史。
