Kaneko H, Hayasaki M, Ohishi I
Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Japan.
Nihon Juigaku Zasshi. 1990 Oct;52(5):995-1000. doi: 10.1292/jvms1939.52.995.
Excretory-secretory (ES) products collected from adult Dirofilaria immitis cultured in vitro were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. ES products of male (M-ES) and female (F-ES) worms were separated into 16 and 21 bands by SDS-PAGE with Coomassie blue and silver staining, respectively. The antigenic bands were then analyzed by immunoblotting, using pooled sera from dogs that had naturally contracted D. immitis. Sera from dogs with microfilaremic infection showed 7 bands in M-ES and 10 bands in F-ES, while those from dogs with occult infection revealed 3 bands in M-ES and 10 bands in F-ES. Among these bands, those of 14, 18, 21, 22, 29, and 32 kilodaltons (Kd) were common to M-ES and F-ES, those of 39 and 44 Kd were specific to M-ES, and those of 20, 38, 43, 53, 63, 90, 110, 125 and 136 Kd were specific to F-ES.
采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹法，对体外培养的成年犬恶丝虫分泌排泄物（ES）产物进行分析。用考马斯亮蓝和银染法，通过SDS-PAGE分别将雄虫（M-ES）和雌虫（F-ES）的ES产物分离成16条和21条带。然后，用自然感染犬恶丝虫的犬混合血清，通过免疫印迹法分析抗原条带。微丝蚴血症感染犬的血清在M-ES中显示7条带，在F-ES中显示10条带；而隐匿感染犬的血清在M-ES中显示3条带，在F-ES中显示10条带。在这些条带中，14、18、21、22、29和32千道尔顿（Kd）的条带是M-ES和F-ES共有的，39和44 Kd的条带是M-ES特有的，20、38、43、53、63、90、110、125和136 Kd的条带是F-ES特有的。
