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자료유형
학술저널
저자정보
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대한바이러스학회 JOURNAL OF BACTERIOLOGY AND VIROLOGY 大韓바이러스學會誌 제18권 제2호
발행연도
1988.12
수록면
149 - 161 (13page)

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The new method for formation of plaques of Hantaviruses was developed using the system of enzyme-labelled antibody and Vero-E6cells maintained with semi-solid rnedium. To establish the method, several preliminary tests were carried out and consecutive studies were included for evaluation and practical application. The obtained conclusions are as follows; 1. The method, immunoenzyme plaque assay, was constructed with 1) anti-Hantavirus antibody and peroxidase labelled anti-rat IgG for antisera, 2) 3, 3-diaminobenzidine tetrahydrochloride for substrate and 3) Vero-E6 monolayer maintained with semisolid methyl cellulose DMEM for virus propagation. 2. The optimal units of antibody and enzyme-labelled antibody were determined as 4-B unit in both. Otherwise, at the higher or lower dilutions, it was not satisfiable in the contrast with background and the intensity of plaque staining. 3. The optimal concentrations of methyl cellulose in culture medium to form the countable, well sized plaques were 0.4% and 0.6% for prospect Hill virus and Hantaan viurs, respectively. 4. The plaques formed by this mathod were proved as to be produced by at least one virus particle according to the result of virus dilution experiment. Also, the observation of plaque by light microscope revealed that the plaque is the group of cells bearing viral antigens which was deposited with reaction product produced histochemically. 5. To determine whether plaques are formed by Hantaan virus, the virus was mixed with either sera from the patients of KHF or sera of rats irnmunized with Hantaan virus. The positive neutralization was evident by remarked reduction of nurnbers of plaques. 6. The sensitivity, specificity and usefulness of the rnethod could be proved by being applied in differential diagnosis of HFRS, analysis of Hantaviral antigens, test for virus interference and massive screening of neutralizing antibody or virus titration.

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