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KMID : 0377619740260050551
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Korean Jungang Medical Journal
1974 Volume.26 No. 5 p.551 ~ p.557
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Antigenic Analysis of an Etiologic Agent of Epidemic Hemorrhagic Conjunetivitis Isolated During 1972 in Korea
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이종훈/Lee, Chong Hoon
이연태/황기선/최상규/고광균/이윤일/면지선/Lee, Yun Tai/Whang, Kee Sun/Choi, Sang Kyu/Ko, Kwang Kjune/Lee, Yoon Il/Byoun, Ji Soun
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Abstract
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An unusual type of conjunctivitis was prevailing in an epidemic fashion in Korea during 1971 and 1972. The conjunctivitis was characterized by severe subconjunctival hemorrhage, lid-swelling and tearing. Its clinical course, however,. was usually shorter than the ordinary epidemic keratoconjunctivitis and completely recovered within 2 weeks after the onset. This disease seemed to´ be highly contagious in unhygienic and crowded public places.
An attempt was made to isolate the causative agent of the epidemic hemorrhagic conjunctivitis by HeLa and LLCMK2 cell cultures. The cytopathic agent was filtered through millipore membrane filters to get rough estimates of the size.
Nucleic acid type, and ethyl ether sensitivity and acid sensitivity were also examined. Sections of LLCMK2 cells infected with the isolated agent were stained with uranylacetate and lead citrate and subjected to electron microscopy to determine the intracellular position and structure of the agents. Cross-neutralization test was done for comparison of antigenic similarity between the isolates and YC-71648 versus with their immune guinea pig sera. In addition the neutralization test was. carried out on 41 sera 6 months after complete recovery from the epidemic hemorrhagic conjunctivitis, 77 healthy sera. The amount of 100 TCID50 of virus was Tested´ against 2 fold-serial-dilution of these sera. Virus serum mixtures Were
incubated at 37°C for 60 minutes and 0.2 ml of the each mixtures was inoculated into 5 LLCMK2 tube cultures. The inoculated cell cultures were adsorbed at 37°C for 2 hours and thereafter kept at 32°C, and a cytopathic effect was observed daily for a week.
The results obtained were as follows:
Two strains of cytopathic agents were isolated by LLCMK2 cell cultures out of 7 conjunctival swabs taken within 2 days after the onset from patients with the -epidemic hemorr hagic conjunctivitis in an outbreak´ 1972.
At the first to third passages by LLCMKZ cell cultures, the cytopathic effect-was often indistinct on the cell monolayer, but it became more clear cut on the 3rd or 4th day of cultivation at the 4th passage, whereas the cytopathic effect was not distinct even after repeated passages on HeLa cell cultures.
Two strains of the isolates could have been gradually adapted to LLCMK2 by passages, as the infectivity titer increased up to 10´-7. ´ TCIDSo per ml at 9th to 10th passages from the titer below 104´° TCID$° per ml until the 8th passages.
The cytopathic agent was very small particle which passed through a membrane filter of pore size 50nm, and spherical in form by elects onphotomicrography.
It was resistant to ethyl ether and acid (pH 3.0) treatment, and the infectivity titer did not drop in the presence of 10-4-°´ Mole per ml of 5-iododeoxyur dine in the culture.
There was a definite cross-neutralization between the YC-71-648 (virus strain isolated in. Japan) and the two isolates with their immune guinea pig sera. A -definite rise in neutralizing antibody titer against the two isolates as well as .Japan strain, was observed on 41 sera obtained from persons 6 months after recovery from the epidemic hemorrhagic conjunctivitis. ´On the other hand, the neutralizing antibody was not detected on 65-66 (84.5-´85.7%) out of 77 healthy sera, except 11-12 sera (14.315.5%) responded to 1 : 4 serum dilution.
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