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Journal of Clinical Microbiology, February 2005, p. 620-628, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.620-628.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Production of Herpes B Virus Recombinant Glycoproteins and Evaluation of Their Diagnostic Potential

Ludmila Perelygina,^* Irina Patrusheva, Soumya Hombaiah, Holley Zurkuhlen, Martin J. Wildes, Nikolai Patrushev, and Julia Hilliard

Viral Immunology Center, Georgia State University, Atlanta, Georgia

Received 29 July 2004/ Returned for modification 29 September 2004/ Accepted 22 October 2004

B virus (cercopithecine herpesvirus 1) is the only deadly alphaherpesvirus that is zoonotically transmissible from macaques to humans. The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals. We evaluated the diagnostic potential of recombinant B virus glycoproteins for the detection of immunoglobulin G (IgG) antibodies in monkey and human sera. Glycoproteins B, C, and E and secreted (sgG) and membrane-associated (mgG) segments of glycoprotein G (gG) were expressed in the baculovirus expression system, while gD was expressed in CHO cells. We developed recombinant protein-based IgG enzyme-linked immunosorbent assays (ELISAs) and compared their diagnostic efficacies by using B virus antibody-negative (n = 40) and -positive (n = 75) macaque sera identified by a whole antigen-based ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG- and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys cross-reacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)- and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data indicate that gB, gC, gD, and mgG have a high diagnostic potential for B virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B virus and those induced by other, closely related alphaherpesviruses, including HSV-1 and -2.

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* Corresponding author. Mailing address: Viral Immunology Center, Georgia State University, P.O. Box 4118, Atlanta, GA 30302-4118. Phone: (404) 651-0002. Fax: (404) 651-0821. E-mail: biolmp{at}panther.gsu.edu.

Present address: Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA 30322.

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Journal of Clinical Microbiology, February 2005, p. 620-628, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.620-628.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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