• Title/Summary/Keyword: western blot/EITB

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Demonstration of species-specific and cross reactive components of Paragonimus tvestermani crude worm antigen by EITB (Immunoblot를 이용한 폐흡충 비항원의 특이 항원대의 증명)

  • Joo, Kyoung-Hwan;Ahn, Hyuck;Chung, Myung-Sook;Lim, Han-Jong
    • Parasites, Hosts and Diseases
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    • v.27 no.1
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    • pp.9-14
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    • 1989
  • Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

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