Gel 여과한 아니사키스 유충항원의 Immunoblot 분석

초록

Anisakiasis is an acute or a chronic gastrointestinal disease caused by the invasion of Anisakis simplex. Nematode larvae of Pseudoterranova decipiens can also infect man. As consumption of raw fish is spread world-widely, anisakiasis becomes now almost a cosmopolitan disease. Acute gastric anisakiasis is rather easily diagnosed by endoscopy or radiological technique. But, preoperational diagnosis of acute intestinal anisakiasis is still difficult because of introducing the gastrofiberscope into the lesion is almost impossible and their clinical manifestations are ambiguous. Furthermore, the diagnosis of chronic anisakiasis is more difficult because the worm invade the stomach or intestinal wall and finally to degenerate. Serodiagnosis may helpful in such diagnostic difficulties. However, any of the serodiagnostic method in anisakiasis has limitations for practical use due to cross reactions. In this context, the study was performed to observe the specifically reacting antigenic bands of gel filtration antigen of the crude extract Anisakis larval worm and their reaction to the serum IgG and IgM antibody obtained from experimentally immunized and infected rabbits using ELISA and SDS-PAGE/immunoblot system. The results obtained were as follows: 1. Using gel filtration, four fractions were obtained (F1 - F4). Protein contents were 0.447 mg/ml, 0.877 mg/ml, 1.240 mg/ml and 0.944 mg/ml respectively. 2. Serum leveles of IgG antibody by ELISA using crude antigen and fractionated antigen except F4 increased from 7 - 9 day and reached their maximum value at 12 - 15 day after infection, and decreased gradually thereafter. Levels of specific IgM antibody elevated from 5-7 day and reached their maximum at 12 day after infection, and decreased gradually thereafter. 3. SDS-PAGE profile showed that each antigen consisted of more than 20 polypeptides of molecular mass 33 - 220 kDa in F1, 27.7 - 220 kDa in F2, 13 - 58 kDa in F3 and below 20 kDa in F4. 4. Using crude antigen and F1, ELISA results was better than other fractions. 5. Major antigenic bands which reacted to specific IgG antibody infected with Anisakis larvae was 46.8 kDa band in each antigens except F1. IgM antibody mainly reacted to the band of 46.8 kDa. Although F1 fraction could be used for ELISA, the low protein content makes it difficult to use in the immunoblot diagnosis of anisakiasis. So, F2 fraction can be considered to be more powerful diagnostic fraction.