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KMID : 0357319930280010055
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Journal of the Korean Society for Microbiology
1993 Volume.28 No. 1 p.55 ~ p.68
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Characteristics of the Human Cytomegalovirus(HCMV) Antigens Binding to Monoclonal Antibody(MCMVA-57, MCMVA-93) and Their Detection by Direct Immunoperoxidase Staining
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김서정
국윤호/박정규/서주영/황응수/박순미/윤종구/차창룡/김길영
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Abstract
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Peroxidase conjugated monoclonal antibodies(MCMVA-57), MCMVA-93) against human cytomegalovirus (HCMV, AD-169) were used to find out characteristics of their antigens and evaluate applicability for diagnostic use.
Molecular weight of their antigens were determined to be 50Kd, 44Kd(MCMVA-57) and 150Kd, 37Kd, 27Kd(MCMVA093). Those antibodies could not neutralize the infectivity of HCMV. By the treatment of several metabolic inhibitors(Cycloheximide,
Actinomycin-D,
Phosphonoacetic acid, Cytosinearabinoside, Tunicamycin), which can differentiate the stages of expression of HCMV antigens, corresponding antigens were concluded to be late proteins and antigens for MCMVA-57 were glycosylated. MCMVA-57 reacted to
three
different kinds of viral particles separated by potassium tartrate gradient ultracentrifugation. MCMVA-93 did not react with any of those viral particles. But they were positive with viral stock solutions prepared by repeated freezing and
thawing.
We
speculated that MCMVA-57 is reactive to one of the glycoproteins on the viral envelope and MCMVA-93 is reactive to one of the precursor of the capsid protein or protein related to DNA. Acetone, xylol, CCL4 are proper fixatives for direct
immunoperoxidase staining.
With those peroxidase conjugated monoclonal antibodies we could detect infected cells on the cell cultures inoculated with patient's urine and in the autopsy specimens(lung) of AIDS patient effeciently.
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KEYWORD
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Human cytomegalovirus antigen, Monoclonal antibody, Direct immunoperoxidase
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