肝吸蟲 感染家兎의 免疫診斷에 對한 硏究 1. 全蟲體 抽出物 粗抗原에 대한 ELISA = Studies on the Immunodiagnosis of Rabbit Clonorchiasis 1. ELISA by whole worm extract crude antigen

다국어 초록 (Multilingual Abstract)

Enzyme-linked immunosorbent a assay(ELIST) using crude antigen of Clonorchis sinensis was performed for infected rabbit sera with different worm burden, from preinfection to 32th or 50th week after infection.
The crude antigen was used the supernatant of homogenated adult worms centrifuged for 1 hr. at 15,000rpm. All rabbits(18) were divided into 4 groups(G-Ⅰ, G-Ⅱ, G-Ⅲ, G-Ⅳ) by different worm burden. The mean of recovered worms(±SD) and the number of rabbits in each group were as follows : G-Ⅰ, 5.4(±2.5) and five : G-Ⅱ, 168.3(±27.4) and three : G-Ⅲ, 317.8(±53.2) and five : G-Ⅳ, 817(±155.6) and five rabbits.
The conjugate used in ELISA was Alkaline phosphatase conjugated anti-rabbit IgG, and substrate was p-nitrophenyl phosphate.
The results were summarized as follows :
1. The optical density(OD) values(antibody levels) as mean of each group increased significantly at the 4th week(P<0.01) in G-Ⅰ : 3rd week (P<0.02) in G-Ⅱ and 4rd week in G-Ⅲ ＆ G-Ⅳ. The mean OD values of groups reached 1.0 or more at 5th week in G-Ⅰ＆ G-Ⅱ and 4rd week in G-Ⅲ ＆ G-Ⅳ. The mean OD values of groups increased every weekly to 10th(G-Ⅱ)-12th(G-Ⅰ ＆ G-Ⅲ)-14th(G-Ⅳ) week, and after that the mean OD values of groups maintained plateau though repeatedly rise and fall(but no significantly) of mean OD values. The mean OD values of each group during plateau period were 1.606-1.794 in G-Ⅰ : 1.719-1.916 in G-Ⅱ : 1.783-1.947 in G-Ⅲ : 1.829->2 in G-Ⅳ.
2. The proportional relationship of worm burden and antibody levels(OD values) were more sensitive until 6th week after infection. The mean OD values of G-Ⅰ were significantly lower than thats of lst week in G-Ⅱ(P<0.02), 3rd-5rd-6th week in G-Ⅲ(P<0.001-P<0.05-P<0.01) and lst-3rd-4rd-5th-6th in G-Ⅳ(P<0.02-P<0.01-P<0.05-p<0.05-P<0.01). The G-Ⅱ were lower than thats of 3rd week in G-Ⅲ ＆ G-Ⅳ(P<0.02), and the G-Ⅲ was lower than that of 3rd week in G-Ⅳ(P<0.05). The mean OD values of all groups were not revealed significant difference(P>0.05) from 8th week to 14th week with each others. The mean OD values of G-Ⅰ sometimes were lower than thats of G-Ⅲ ＆ G-Ⅳ from 16th week to 32th week, and the mean OD values of G-Ⅱ ＆ G-Ⅲ ＆ G-Ⅳ were not revealed significant difference except 3rd week.
3. The individual animals of all groups became OD values 1.0 and more at 3rd to 5th weeks(except one, at 8th). The antibody levels in several individuals were not closely related with worm burden. The OD values were not reached to OD values 1.0 over in all individuals of G-Ⅰ. The individuals in G-Ⅲ ＆ G-Ⅳ were showed OD values 2.0 over(>2) for remaining weeks after 16th week approximately.
Consequently, the detection of antibody of rabbit clonorchiasis in ELISA was very sensitive, but there were a difference of antibody levels according to individual animals.