İNDİREKT ELISA YÖNTEMİNİN TAVUK HASTALIKLARININ TEŞHİSİNDE UYGULANMA OLANAKLARI VE DİĞER TEŞHiS YÖNTEMLERİ İLE KARŞILAŞTIRILMASI

Öz

Son yıllarda soluble antijen veya antikorların enzim immunoassay ile solid faz hale getirilip teşhis edilmesinde büyük gelişmeler kaydedilmiştir. lrrımunoassay, diğer immunokimyasal tekniklerin en önemlilerinden biri olup aynı zamanda diğer tekniklerden daha az komplike, daha kolay ve daha çabuktur (21). Antijen ve antikor tayininde oldukca hassas olan ve ELISA olarak bilinen test 1970'lerin başında Engvall ve arkadaşları Van Weemen ile Schurs tarafından geliştirilmiştir( 18). ELISA yöntemi ile antikorların veya antijenlerin saptanmasında ortam ya antikor yada antijen ile kaplanarak duyarlaştırılır ve üzerine konan antijen veya antikor ile reaksiyona girer, üzerine ilave edilen enzim işaretli antiimmunoglobulin ve substrat ile renk değişimi şeklinde substratın parçalanma oranına göre test sonuçlanır ve mikropleytde oluşturulan bu reaksiyonda bir pleyt üzerinde çok sayıda numune işlenebilmektedir. Bu araştırmada şu anda ülkemiz tavukculuğunun önemli hastalıklardan Newcastle, Gumboro, Epidemik tremor ve Enfeksiyöz bronşitis için antikor tayin etmekte kullanılacak olan indirekt ELI SA metodu laboratuvarıınızda standardize edilmiştir.

Anahtar Kelimeler

• İNDİREKT ELISA
• ND
• IBD
• IB
• AE
• TAVUK HASTALIKLARI

Application of indirect ELISA for the diagnosis of poultry diseases and comparison with the other test methods

Öz

The ELISA assay is an antigen-antibody reaction system which applicates an antibody coupled enzyme with the substrate is directly proportional to the amount of antibody or antigen present in the sample being tested. lmmunassay was the linkage of solu be antigen or antibody to an insoluble soild phase in a way in which reactivity of the iımnunological component was retained. This was the basis of the techniques known ELISA (Enzyme Linked lmmunosorbent Assay) pioneered by Engvall and colleaques Van Weemen and Schuurs (18). With the appropriate assay can be rcmarkably quick and easy yielding infommtion that would be diffıcult to .detennine by the other tecniques (21 ). The ELISA was fırst introduced in the early 1970's andisa highly sensitive method for assaying either antigens or antibodies. In principle the assay is analogus to radio-immunoassay (RIA), the only major difference being to use of an enzyme other that an isotope. Heterogenous enzyme-immunoassays combine the advantages of inumınoflourescense and radioimmunoassay and overcome many of the disadvantages of the other two methods. Enzyme labelled reagents are cheap, easy to prepare and are highly stable so giving a long shelf life, yet yield assays which approach the sensitivity of radionumıassay and which give objective result that can be determined, either visually or with rather simple equipment. In this study, we standardized to determine the antibody with indirect ELISA technique for Gumboro, Newcastle, Avian Encephalomyelitis and lnfectious Bronchitis diseases of the im portant poulty diseases of o ur country. The ELISA and research labotory of our institute will go on to developELJSA test methods for the other important diseases.

Anahtar Kelimeler

• INDIRECT ELISA
• ND
• IBD
• IB
• AE

Kaynakça

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13. SNYDER, D.B., MARQUARDT, W. W., MALLINSON, E. T. and RUSSEK, E.1982 : Rapid Serological Profiling by Enzyme-Linked lmmunorsorbent Assay. I.Measurement of Antibody Activity Titer Againts Newcastle Disease Virus in a single serum diluition. Avian Dis 27:161-170.
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15. SNYDER, D. B.,Marquarty, W. W., MALLINSON, E. T., ALLEN, D. C., 1985 : Rapid Serologyval profiling by Enzme-Linked lmmunosorbent Assay. III. Simultaneous measurements ofantibody titters to ınfectious Bronchitis, Jncfectious Bursal Disease and Newcastle Disease Viruses in a msingle serum dillution. Avian Dis. 28 :12-24
16. SNYEDER, D. B., MARQUARDT. W. W., MALLISON, E. T., and RUSSEKCOHEN- E- SAVAGE, P. K.and ALLEN, D. C. 1985 : Rapid Serologycal Profiling by enzyme-Linked Immunosorbent Assay. IV. Association of infectious Bursal Disease Serology with Broiler Flock Performance. Avian Dis.30: 139-148.
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18. VOL LER, A. 1976 : The application of mi c ro-plate enzyme-linked immunosorbent assays to some infectious disease. Firs't International Symposium on lmmunoenzymatic techniques INSERM Symposium No:2 North/Holland Publishing Compeny Amsterdam. 167-173 .
19. VOLLER, A. 1978 : Workshop report on ELİSA techniques Immunofluorescence and related staining techniques. Elsevier/North Holland Biomedical Press 337-347 .
20. VOLLER, A., BIDWELL; D. E. and BARTLETT, A. 1979 : Types of Assay and the applications ELISA. The Enzyme Linked Immunosorbent Assay (ELİSA) 1. st. Edition, The Zoological Society of London. 3-8. 10-58.
21. WEYBRIDGE Central Veterinary Laboratory, U.K. 1988 : Diagnostic produtes control section method sheets. 3-70.