The Evaluation of Recornbinant Circumsporozoite Protein in Malaria Diagnosis

말라리아 진단시 재조합 Circumsporozoite 단백질의 유용성 평가

  • 이형우 (국립보건원 바이러스질환부 의동물과) ;
  • 이종수 (국립보건원 바이러스질환부 의동물과) ;
  • 이원자 (국립보건원 의동물과) ;
  • 조신형 (국립보건원 바이러스질환부 의동불과) ;
  • 이호자 (경희대학교 생물학과)
  • Published : 2000.06.01

Abstract

It had been evaluated the recombinant Circumsporozoite(CS) protein of Plasmodium viva in serologic diagnosis of vivax malaria. Western blot was done to analyse the sera of malaria patients according to the days after onset. The sera which have the terms within 15 days were shown 43.8%(14/32) of positive rates and the sera over the 16 days were shown 94.4%(17/18) of positive rates. So the total positive rate was 62%(31/50). It was 22.6%(7/31) which was shown negative response in Western blot, even though they were shown positive response in Immuuofluorescent antibody test(1FAT) using whole blood stage antigens. The positive rate of non-epidemic area(Yechon-gun, Kyongsangbuk-do) was 10.7%(3/28), and epidemic area(Kangwha-gun, Inchon-shi) was 27.6%(13/47) in Western blot analysis using recombinant CS protein. In order to applicate the recombinant CS protein in seroepidemiological survey, blood samples of 422 inhabitants were collected who lived in malaria epidemic areas, Chosm-ri, Majeong-ri, Hyangyang-ri and Noejo-n in Paju-shi, Kyonggi-do. All of them were negative in microscopic examination and two(0.5%) of them were positive in Polymerase Chain Reaction. 42(10.0%) of them were seropositive in FAT using whole blood antigens and 71(16.8%) of them were seropositive in Enzyme-linked immunosorbent assay using recombinant CS protein. It was figured out the positive rates were much higher according to the distances of villages which were closed to the demilitalized zone(DMZ) in all kind of diagnostic methods, respectively.

국내에서 유행하는 Plasmodium vivax의 유전자를 조작하여 만든 재조항 Circumsporozrozite(CS) 단백질 (22)을 이용하여 말라리아에 대한 혈청학적 검사의 유용성을 평가하였다. 최초발병일로부터 진단까지 걸린 기간을 기준으로 환자들의 면역반응을 Western blot으로 알아본 결과, 15일 이내에 진단받은 환자들은 43.8%(14/32)가 양성반응을 보였고, 16일 이상 경과한 환자들은 94.4%(17/18)가 양성반응을 보여 전체적으로는 62%(31/50)의 양성률을 보였다. Blood stage 항원에 대한 항체를 갖고 있음에도 불구하고 재조합 CS단백질 항원에 대해 음성인 환자들이 22.6%(7/31)였다. 비유행지 주민(경북 예천군)들은 10.7%(3/28),유행시 주민(인천시 강화군)들은 27.6%(13/47)의 재조합 CS 단백질에 대한 항체 양성을 보였다. 재조합 CS 단백질 항원의 역학조사시 유용성을 알아보기 위하여 말라리아 유행지인 경기도 파주시 조산리, 마정리, 항양리, 뇌조리 거주 주민 422명의 전혈을 채취하여 혈액도말법과 중합효소연쇄 반응법으로 항원검사를 실시하였으며 blood stage 항원을 이용한 간접면역형광법과 재조합 CS 단백질 항원을 이용한 효소면역측정법으로 말라리아 항체보유 여부를 조사하였다. 혈액도말법에서는 422명 모두 음성이었으나, 중합효소연쇄반응법에서는 2명(0.47%)이 양성이었다. 간접면역형광법에서는 42명(9.95%), 재조합 CS 단백질 항원을 이용한 효소변역측정법에서는 71명(16.82%)이 양성으로 나타났다. 두 검사에서 모두 항체 양성을 보인 사람은 8명(11.27%)에 불과하였다.

Keywords

References

  1. 대한수혈학회지 v.8 말라리아 호발지역에서 채혈된 주민과 군헌혈자에서 말라리아 검사성적 임채승;김영기;이갑노;이형우;이원자;이종수;조남선;김대성
  2. 대한감염학회 학술대회초록집 v.120 재조합 Circumsporozoite protein 항원을 이용한 Antibody ELISA의 유용성 평가 임채승;김태성;김민자;조성원;이은일;송진원;송기준;노천섭;김상훈;박승철
  3. 수혈을 통한 말라리아가 원인었던 불명열 1예감염 v.31 정인경;오명돈;채종일;이형우;이원자;이종수;서동희;최강원
  4. 대한의학학회98년도심포지움 국내 말라리아 유행의 전망과 대책 채종일
  5. Esp. Parasitol. v.70 DNA probe diagnisis of parasitic infections Baker, R.H.Jr.
  6. Am. J. Trop. Med. Hyg. v.46 A simple method to detect Plasnmodium falciparum directly from blood samples using the polymerase chain reaction Barker, R.H.Jr.;Banchongaksorn, T.;Courval, J.M.;Suwonked, W.;Rimwungtragoon, K.;Wirth, D.E.
  7. Am. J. Trop. Med. Hyg. v.44 Characteristics of natural antibody responses to the circumsporozoite protein of Plasmodium vivax Brown, A.E.;Webster, H.K.;Krinchal, K. Gordon, D.M.;Wirtz. R. A.;Permpanich, B.
  8. Vivax infections undetected by convertional microscopy. Trans. R. Soc. Trop.Med. v.86 Demonstration by the polymerase chain reaction of mixed Plasmodium falciparum and P Brown, A.E.;Kain, K.C.;Pipithkul, J.;Webster, K.
  9. Am. J. Trop. Med. Hyg. v.25 An evaluation of malaria fluorescent antibody patterns in several remote island populations of the New Guinea Brown, P.;Collins, W.E.;Gajdusek, D.C.;Miller, L.H.
  10. Trans. R. Soc. Trop. Med. Hyg. v.83 Comparison of antibody responses to the circumsporozoite protein repeat region and to intact sporozoites during acute falciparum malaria Brown, A.E.;Webster, H.K.;Pavanand, K. Permpanich, B.;Sookio, P.;Sattabongkot, J.;Gingrich, J.B.
  11. P. vivax circumsporozoite proteins in a human population. J. Clin. Microbiol. v.27 Differential antibody responses to Plasmodium falciparum Burkot, T.R.;Graves,P.M.;Wirtz, R.A.;Brabin, B.J.;Battistutta D.;Cattani, J.A.;Maizels, R.M.Alpers, M.P.
  12. Bull. W. H. O. v.39 Studies on the relationship between fluorescent antibody response and ecology of malaria in Malaysia Collins, W.E.;Warren, M.;Skinner, J.C. Fredericks, H.J.
  13. Am. J. Trop. Med. Hyg. v.21 The indirect fluorescent antibody test for malaria. Collins, W.E.;Skinner, J.C.
  14. V. Response of sera from Nigerians to five Plasmodium antigens. Am. J. Trop. Med. Hyg. v.16 Fluorescent antibody studies on human malaria. Collins, W.E.;Skinner, J.C. Coifman, R.E.
  15. Infect. Immun. v.53 Level of antibodies to Plasmodium falciparum sporozoite surface antigens reflect malaria transmission rates and are persistent in the absence of reinfection Druihe, P.;Pradier, O.;Marc, J.P.;Miltgen, F.;Mazier, D.;Parent,G.
  16. Trans. R. Soc. Trop. Med. Hyg. v.82 Prevalence and levels of antibodies to the circumsporozoite protein of Plasmodium falciparum in an endemic area and their relationship to resistance against malaria infection Esposito, F.;Lombardi, S,;Modiano, D.;Zavala, F.Reeme, J.;Lamizana, L.;Coluzzi, M.;Nussenzweig, R.S.
  17. Am. J. Trop. Med. Hyg. v.44 Plasmodium vivax sporozoite antibodies in individuals exposed during a single malaria outbreak in a non-endemic area Fontes, C.C.J.;Bathurst, I.;Krettli, A.U.
  18. Am. J. Trop. Med. Hyg. v.37 Use of a DNA hybridization assay for the detection of Plasmodium falciparum in field trials Holmberg, M.;Shenton, F.C.;Janneth, K.Snow,R.W.Pettersson, U.;Wigzell, H.; Greenwwod, B.M.
  19. Am, J. Trop. Med. Hyg. v.24 Application of the indirect fluorascent antibody method in a study of malaria endemicity in Mato Grosso Brazil Jeffery, G.M.;Warren, M.;Collins, W.E. Lobel, H.
  20. J. Infec. Dis. v.168 Detection of Plasmodium vivax by polymerase chain reaction in a field study Kain, K.C.;Brown, A.E.;Mirabelli, L.;Webster, H.K.
  21. Am. J. Trop. Med. Hyg. v.50 Use or the polymerase chain reaction to directly detect malaria parasites in blood samples from the Venezuelan Amazon Laserson, K.F.;Petralanda, I.;Hamilin, D.M.;Almera, R.;Fuentes, M.;Carrasquel, A;Barker Jr, R.H.
  22. Kor. J. Microbiol. v.37 DNA sequencing and expression of the circumsporozotie protein of Plasmodium vivax Korean isolate in Escherichia coli Lee, H.W.;Lee, J.S.;Lee, W.J.;Lee, H.S.
  23. Korea. Kor. J. Parasitol v.36 Current status of vivax malaria among civilians in Korea Lee, J.S.;Kho, W.G.;Lee, H.W.;Seo, M.;Lee, W,J.
  24. Am. J. Thop. Med. Hyg. v.34 Detection of Plasmodium falciparum using a synthetic DNA probe Mclaughlin, G.L.;Edlind, T.D.;Campbell, G.H.;Eller, R.F.;Ihler, G.M.
  25. Parasit, Immunol. v.14 Anti-sporozoite protein antibodies measure age related exposure to malaria in Kataragama Sri Lanka Mendis, C.;Giudice,G.D.;Mendis, A.C.G.;Tougne.C.;pessi, A.;Weerasinghe, S.;Carter, R.;Mendis, K.N.
  26. Exp. Parasit. v.76 Plasmodium vivax;Recombination between potential allelic types of the merozoite surface protein MSP-1 in parasites isolated from patients Premawansa,S.;Snewin, V. A.;Khouri, E.;Mendis,K.N.;David, P.H.
  27. Vaccine. v.14 Production of recombinant SERA proteins of Plasmodium falciparum in Escherichia coli by using synthetic genes Sugiyama, T.;Suzue, K.;Okamoto, M.;Inselburg, J.;Tai, K.;Horii,T.
  28. Am. J. Trop. Med. Hyg. v.26 Malaria endemicity among Orang Asli(Malasian aborigines)as detemined by indirect fluorescent antibody tests Thomas, V.; Dissanaike, A.S.
  29. Am. J. Trop. Med. Hyg. v.51 A highly sensitive rapid and simple polymerase chain reaction-based method to detect human malaria(Plasmodium falciparum and Plasmodium vivax) in blood samples Tirasophon, W.;Rajkulchai, P.;Ponglikitmongkol. M.;Wilairat,P.;Boonsaeng, V.;Panyim, S.
  30. Am. Trop. Med. Hyg. v.46 Application of seroepidemiology in identification of local foci in a malarious community in Palawan The Philippines Tongol-Rivera, P.;Kano, S.;Migule, E.;Suzuki, M.
  31. Method. in Enzymol. v.92 Enzymelinked immunoelectrotransfer blot techniques(EITB) for studying the specificities of antigens and antibodies separated by gel electrophoreses Tsang, V.C.W.;Peralta, J.M. Simons, A.R.
  32. Proc. Soc. Exp. Biol.med. v.110 Fluorescent antibody staining as a measure of malarial antibody Voller, A.; Bray, R.S.
  33. J. Ciln. Microbiol. v.25 Development of immunity in natural plasmodium falciparum malaria:antibodies to the falciparum sporozoite vaccine I antigen(R32tet32) Webster, H.K.;Boudreau,E.F.;Pang, L.W.;Permpanich, B.;Sookto, P.;Writz, R.A.
  34. Am. J. Trop. Med. Hyg. v.47 Circumsporozoite antibody as a serologic marked of plasmodium falciparum transmission Webster, H.K.;Gingrich, J.B.;Wongsrichanalai, C.;Tulyayon, S.;Suvarnamani, A.; Sookto, P.;Permpanich, B.
  35. Trans. R. Soc. Trop. Med. Hyg. v.84 Antibodies to plasmodium falciparum soporozoites following a malarial outbreak in a nonendemic area of Sri Lanka Wijesundera, M.;de, S,;peiris, J.S.M.;Ariyaratne, Y.G.;Verdini, A.S.;Pessi, A.;Giudice, G.D.
  36. Exp. parasitol. v.63 Plasmodium falciparum: immunogenicity of circumsporozoite protein constructs produced in Escherichia coli. Wirtz, R.A.;Ballou, W.R.;Schneider, I, Chedid, L, Gross, M.J.;Yong, J.F.;Hollingdale, M.;Diggs, C.L.;Hockmeyer, W.T.
  37. Exp. parasitol. v.75 Plasmodium falciparum: a simple polymerase chain reaction method for differentiating strains Wooden, J.;Gould,E.E.;Paull,A.T.;Sibley, C.H.
  38. Scinece. v.228 Expression of Plasmodium falciparum circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine Young, J.F.;Hockmeyer,W.T.;Gross, M.;Ballou, W.R.;Wirtz, R.A,;Troper, J.H.;Beaudoin, R.l.;Hollingdale, M.R.;Miller,L.H.;Diggs, C.L.;Rogenberg, M.
  39. Science v.228 Expression of Plasmodium falciparum circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine Young,J.F.;W.T.Hockmeyer;M.Gross;W.R.Ballou;R.A.Wirtz;J.H.Troper;R.L.Beaudoin;M.R.Hollingdale;L.H.Miller;C.L.Diggs;M.Rogenberg