Öz
The aims of this study were to determine the antigenic differences of hydatid cyst fluid from different animals and to investigate the effect of the origin of antigenic materials on the diagnosis of hydatidosis in humans and mice. For this purpose, the proteins of hydatid cyst fluid (HCF) isolated from the liver of sheep and donkeys and secondary hydatid cysts fluid (SHCF) from albino mice infected with protoscolices isolated from sheep and donkeys were separated by SDS-PAGE. This study was performed at the University of Ankara, Faculty of Veterinary Medicine, Department of Parasitology. For comparing antigenic profiles and detecting specific protein bands of those antigens, 10 infected and 10 non-infected human sera were tested for cystic echinococcosis (CE), and 16 positive and 9 negative mice sera were tested for secondary hydatidosis (SH) by using Western blotting technique. The specific protein band for mice sera was detected as 8 kDa (100%) for sheep’s and donkey’s hydatid cyst fluid. The specific proteins for human sera were detected at 24 kDa (100%) and at 8 kDa (100%) for sheep’s and donkey’s hydatid cyst fluid respectively. No band was detected in negative mice and human sera. It was concluded that SHCF antigen from mice could be considered useful as being easy to provide and give good results for the diagnosis of CE
Anahtar Kelimeler
Cyst fluid antigen diagnosis donkey mice sheep Western blotting
Kaynakça
- Barıs I, Sahin A, Bilir N, Kalyoncu AF, Emri AS, Akhan O, Barıs B, Copur AS, Selcuk ZT (1998): Hidatik kist hastalığı ve Türkiye' deki konumu. Türkiye Akciğer Hastalıkları Vakfı Yayını, No:1, Ankara.
- Craig PS, Rogan MT, Allan JC (1996): Detection, screening and community epidemiology of taeniid cestode zoonosis: cystic echinococcosis, alveolar echinococcosis and neurocysticercosis. Adv Parasit, 38,169–250.
- Gonzales G, Nieto A, Fernandez C, Orn A, Wernstedt C, Hellman U (1996): Two different 8 kDa monomers are involved in the oligomeric organization of the native Echinococcus granulosus antigen B. Parasite Immunol, 18, 587–596.
- Güralp N (1981): Helmintoloji. Ankara Üniv. Vet. Fak. Yayınları, No: 368, Ankara.
- Ioppolo S, Notargiacomo S, Profumo E, Frenchi C, Ortona E, Rigano R, Siracusano A (1996): Immunological responses to antigen B from Echinococcus granulosus cyst fluid in hydatid patients. Parasite Immunol, 18, 571– 578.
- Irabuena O, Nieto A, Ferreira AM, Battistoni J, Ferragut G (2000): Characterization and optimization of bovine Echinococcus granulosus cyst fluid to be used in immunodiagnosis of hydatid disease by ELISA. Rev Inst Med Trop Sao Paulo, 42, 255–262.
- Ito A, Ma L, Schantz PM, Gottstein B, Liu YH, Chai JJ, Abdelhafez SK, Altıntas N, Joshi DD, Lightowlers MW, Pawlowski ZS (1999): Differential serodiagnosis for cystic and alveolar echinococcosis using fractions of Echinococcus granulosus cyst fluid (antigen B) and Echinococcus multilocularis protoscolex (EM18). Am J Trop Med Hyg, 60, 188–192.
- Ito A, Okamoto M, Ishiguro T, Ma L, Suzuki H, Yasui A, Shigeta H, Matsuura T, Hosokawa T, Chai TT (1998): An imported case of cystic echinococcosis in Japan diagnosed by imaging and serology with confirmation of Echinococcus granulosus-specific DNA sequences. Am J Trop Med Hyg, 58, 790–792.
- Laemmli UK (1970): Cleava of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680.
- Leggatt GR, Yang W, Mcmanus DP (1992): Serological evaluation of the 12 kDa subunit of antigen B in Echinococcus granulosus cyst fluid by immunoblot analysis. T Roy Soc Trop Med H, 86, 189–192.
- Lightowlers M, Gottstein B (1995): Immunodiagnosis of echinococcosis. 355-410. In: RCA Thompson, AJ Lymbery (Eds), Echinococcus and Hydatid Disease. CAB International, Wallingford, United Kingdom.
- Mamuti W, Yamasaki H, Sako Y, Nakaya K, Nakao M, Lightowlers MW, Ito A (2002): Usefulness of hydatid cyst fluid of Echinococcus granulosus developed in mice with secondary infection for serodiagnosis of cystic Echinococcosis in humans. Clin Diagn Lab Immunol, 9, 573-576.
- Margutti P, Ortona E, Vaccari S, Barca S, Rigano R, Teggi A, Muhschlegel F, Frosch M, Siracusano A (1999): Cloning and expression of a cDNA encoding an elongation factor 1beta/delta protein from Echinococcus granulosus with immunogenic activity. Parasite Immunol, 21, 485–492.
- Meddison SE, Selemanda SB, Schantz PM, Fried JA, Wilson M, Tsang VCW (1989): A specific diagnostic antigen of Echinococcus granulosus with an apparent molecular weight of 8 kDa. Am J Trop Med Hyg, 40, 377– 383.
- Poretti D, Felleisen E, Grimm F, Pfister M, Teuscher F, Zuercher C, Reichen J, Gottstein B (1999): Differential immunodiagnosis between cystic hydatid disease and other cross-reactive pathologies. Am J Trop Med Hyg, 60, 193– 198.
- Sarkari B, Sadjjadi SM, Abidi H, Izadpanah A,
- Kazemian S, Rafati A (2007): Application of Western Blotting Using Native Antigen B for Serodiagnosis of Human Cystic Echinococcosis. Iran J Parasitol, 2, 7-12.
- Schantz PM, Chai J, Craig PS, Eckert J, Jenkins DJ, Macpherson CNL, Thakur A (1995): Epidemiology and control of hydatid disease. 233-331. In: RCA Thompson, AJ Lymbery (Eds), Echinococcus and Hydatid Disease. CAB International, Wallingford, United Kingdom.
- Sharma SD, Mullenax J, Araujo FG (1987): Western blot analysis of the antigens of T. gondii recognized by human IgM antibodies. J Immunol, 131, 977-978.
- Siracuso A, Ioppolo S, Notargiacomo S, Ortona E, Rigano R, Teggi A, Derosa F, Vicari G (1991): Detection of antibodies against Echinococcus granulosus major antigens and their subunits by immunoblotting. T Roy Soc Trop Med H, 85, 239–243.
- Towbin H, Staehelin T, Gordon J (1979): Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. P Natl Acad Sci USA, 76, 4350
Öz
Çalışmanın amacı farklı hayvanların hidatik kist sıvısının antijenik farklılıklarını saptamak ve insan ve farelerde hidatidosisin identifikasyonunda antijenik materyalin orijininin etkisini araştırmaktır. Bu amaçla Ankara Üniversitesi Veteriner Fakültesi Parazitoloji Anabilim Dalında yapılan çalışmada, koyun ve eşek karaciğerlerinden elde edilen hidatik kist sıvılarındaki proteinler ile yine koyun ve eşek orijinli protoskolekslerle enfekte albino farelerden elde edilen sekonder hidatik kist sıvılarındaki proteinler SDS-PAGE yöntemi ile ayrılma işlemine tabi tutulmuştur. Bu antijenik yapıların spesifik protein bantlarının tespiti ve karşılaştırılması için kistik ekinokokoz yönünden 10 enfekte, 10 enfekte olmayan insan serumu ve sekonder hidatidosis için 16 pozitif ve 9 negatif fare serumu Western blot yöntemi ile test edilmiştir. Fare serumlarında koyun ve eşek kist sıvıları için tek spesifik protein bandının 8 kDa (%100) olduğu saptanırken, insan serumları kullanıldığında koyun hidatik kist sıvısı için 24 kDa (%100), eşek hidatik kist sıvısı için 8 kDa (%100) bandın spesifik olduğu tespit edilmiştir. İnsan ve fare negatif serumlarında banda rastlanmamıştır. Sonuç olarak kistik ekinikokozun teşhisinde farelerden elde edilen sekonder hidatik kist sıvılarının iyi sonuç verdiği ve elde edilmesinin kolay olması yönüyle hastalığın tanısında rahatça kullanılacağı ortaya konmuştur
Anahtar Kelimeler
Kaynakça
- Barıs I, Sahin A, Bilir N, Kalyoncu AF, Emri AS, Akhan O, Barıs B, Copur AS, Selcuk ZT (1998): Hidatik kist hastalığı ve Türkiye' deki konumu. Türkiye Akciğer Hastalıkları Vakfı Yayını, No:1, Ankara.
- Craig PS, Rogan MT, Allan JC (1996): Detection, screening and community epidemiology of taeniid cestode zoonosis: cystic echinococcosis, alveolar echinococcosis and neurocysticercosis. Adv Parasit, 38,169–250.
- Gonzales G, Nieto A, Fernandez C, Orn A, Wernstedt C, Hellman U (1996): Two different 8 kDa monomers are involved in the oligomeric organization of the native Echinococcus granulosus antigen B. Parasite Immunol, 18, 587–596.
- Güralp N (1981): Helmintoloji. Ankara Üniv. Vet. Fak. Yayınları, No: 368, Ankara.
- Ioppolo S, Notargiacomo S, Profumo E, Frenchi C, Ortona E, Rigano R, Siracusano A (1996): Immunological responses to antigen B from Echinococcus granulosus cyst fluid in hydatid patients. Parasite Immunol, 18, 571– 578.
- Irabuena O, Nieto A, Ferreira AM, Battistoni J, Ferragut G (2000): Characterization and optimization of bovine Echinococcus granulosus cyst fluid to be used in immunodiagnosis of hydatid disease by ELISA. Rev Inst Med Trop Sao Paulo, 42, 255–262.
- Ito A, Ma L, Schantz PM, Gottstein B, Liu YH, Chai JJ, Abdelhafez SK, Altıntas N, Joshi DD, Lightowlers MW, Pawlowski ZS (1999): Differential serodiagnosis for cystic and alveolar echinococcosis using fractions of Echinococcus granulosus cyst fluid (antigen B) and Echinococcus multilocularis protoscolex (EM18). Am J Trop Med Hyg, 60, 188–192.
- Ito A, Okamoto M, Ishiguro T, Ma L, Suzuki H, Yasui A, Shigeta H, Matsuura T, Hosokawa T, Chai TT (1998): An imported case of cystic echinococcosis in Japan diagnosed by imaging and serology with confirmation of Echinococcus granulosus-specific DNA sequences. Am J Trop Med Hyg, 58, 790–792.
- Laemmli UK (1970): Cleava of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680.
- Leggatt GR, Yang W, Mcmanus DP (1992): Serological evaluation of the 12 kDa subunit of antigen B in Echinococcus granulosus cyst fluid by immunoblot analysis. T Roy Soc Trop Med H, 86, 189–192.
- Lightowlers M, Gottstein B (1995): Immunodiagnosis of echinococcosis. 355-410. In: RCA Thompson, AJ Lymbery (Eds), Echinococcus and Hydatid Disease. CAB International, Wallingford, United Kingdom.
- Mamuti W, Yamasaki H, Sako Y, Nakaya K, Nakao M, Lightowlers MW, Ito A (2002): Usefulness of hydatid cyst fluid of Echinococcus granulosus developed in mice with secondary infection for serodiagnosis of cystic Echinococcosis in humans. Clin Diagn Lab Immunol, 9, 573-576.
- Margutti P, Ortona E, Vaccari S, Barca S, Rigano R, Teggi A, Muhschlegel F, Frosch M, Siracusano A (1999): Cloning and expression of a cDNA encoding an elongation factor 1beta/delta protein from Echinococcus granulosus with immunogenic activity. Parasite Immunol, 21, 485–492.
- Meddison SE, Selemanda SB, Schantz PM, Fried JA, Wilson M, Tsang VCW (1989): A specific diagnostic antigen of Echinococcus granulosus with an apparent molecular weight of 8 kDa. Am J Trop Med Hyg, 40, 377– 383.
- Poretti D, Felleisen E, Grimm F, Pfister M, Teuscher F, Zuercher C, Reichen J, Gottstein B (1999): Differential immunodiagnosis between cystic hydatid disease and other cross-reactive pathologies. Am J Trop Med Hyg, 60, 193– 198.
- Sarkari B, Sadjjadi SM, Abidi H, Izadpanah A,
- Kazemian S, Rafati A (2007): Application of Western Blotting Using Native Antigen B for Serodiagnosis of Human Cystic Echinococcosis. Iran J Parasitol, 2, 7-12.
- Schantz PM, Chai J, Craig PS, Eckert J, Jenkins DJ, Macpherson CNL, Thakur A (1995): Epidemiology and control of hydatid disease. 233-331. In: RCA Thompson, AJ Lymbery (Eds), Echinococcus and Hydatid Disease. CAB International, Wallingford, United Kingdom.
- Sharma SD, Mullenax J, Araujo FG (1987): Western blot analysis of the antigens of T. gondii recognized by human IgM antibodies. J Immunol, 131, 977-978.
- Siracuso A, Ioppolo S, Notargiacomo S, Ortona E, Rigano R, Teggi A, Derosa F, Vicari G (1991): Detection of antibodies against Echinococcus granulosus major antigens and their subunits by immunoblotting. T Roy Soc Trop Med H, 85, 239–243.
- Towbin H, Staehelin T, Gordon J (1979): Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. P Natl Acad Sci USA, 76, 4350
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Veteriner Cerrahi |
| Diğer ID | JA86EV88SD |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 1 Mart 2013 |
| Yayımlandığı Sayı | Yıl 2013 |