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KMID : 0357319920270030289
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Journal of the Korean Society for Microbiology
1992 Volume.27 No. 3 p.289 ~ p.296
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Quantiation of Human Cytomegalovirus by Dot-blot Immunoassay
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황응수
박순미/박정규/균윤호/김익상/최성배/장우현/차창룡
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Abstract
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A rapid and simple dot blot immuoassay using monoclonal antibody was developed which can be used directly to identify and quantitate human cytomegalovirus (HCMV) directly in urine sample without cell culture.
Concentrated urine samples were serially diluted and blotted onto a strip of nitrocellulose membrane. The strip was stained immunoenzymatically with MCMVA66 which recognized cytoplasmic component of HCMV-infected cells. The quantity of HCMV was
expressed as dot blot unit (DBU, reciprocal of endpoint titer of dot blot immunoassay) and was compared with conventional quantitative unit such as ICU (infected cell unit) which represented the number of immunoezymatically stained HCMV-infected
cells
and PFU (plaque forming unit). Conversion of DBV into ICU or PFU showed linear correlation between DBU and conventional quantitative unit. The result was as follows:66 DBU:312.5 ICU/0.2ml or 40.6 PFU/0.2ml, 8 DBU:25 0ICU/0.2ml or 32.5 PFU/0.2ml,
and 1
DBU:200 ICU/0.2ml or 26 PFU/0.2ml. This assay therefore could be a useful method for rapid (under the optimal working conditions it took about 9 hrs to get the result), sensitive and acourate quantitation of HCMV, especially for the viruses
excreted in
clinical samples. It also would be useful for assessing the effect of antiviral drugs in vivo.
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KEYWORD
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