Öz
In this study, use and standardization of immunofluorescence technique for the diagnosis of infectious bursal disease (IBD) infection and preparation of the conjugate that will be used in the technique were aimed. Immunized chicken and rabbit sera were labelled with fluorescein isothiocyanate. In the standardization of the conjugate dilution, optimal ratio was determined as 1:16 after the evaluation of smears prepared from the bursa Fabricius of the control and experimentally infected groups. Indirect fluorescent antibody technique was found to be relatively superior to the direct fluorescent antibody technique after the standardization tests. In the field studies, 23 out of 28 (85.13%) IBDV infected broiler flocks were detected to be positive by immunofluorescence technique. As a conclusion, immunofluorescent antibody technique (IFAT) was found to be useful for diagnosis of the IBDV infections
Anahtar Kelimeler
Kaynakça
- Akan M (2002): İnfeksiyöz bursal hastalık. 169-178. In: Kanatlı Hayvan Hastalıkları. İzgür M, Akan M. (Eds), Medisan Yayınevi, Ankara.
- Allen, GM, McNulty MS, Connor TJ, McCracken RM, McFerran JB (1984): Rapid diagnosis of infectious bursal disease infection by immunoflourescence on clinical material. Avian Pathol, 13, 419-427.
- Anonim (1996): Infectious bursal disease (Gumboro disease). Chapter 3. 6. 1. In: Manual of Standarts for Diagnostic Tests and Vaccines. OIE Lists A and B diseases of mammals, birds and bees. 3th edition, Paris. An immunoperoxidase monoclonal antibody stain for rapid diagnosis of infectious bursal disease. Avian Dis, 31, 538- 545.
- Cruz-Coy JS, Giambrone JJ, Panangala VS (1993): Production and characterization of monoclonal antibodies against variant A infectious bursal disease virus. Avian Dis, 37, 406-411.
- Goldman M, Carver RK (1957): Preserving flourescein isocyanate for simplified preparation of fluorescent antibody. Science, 126, 839-840.
- Goldman M (1968): Fluorescent Antibody Methods. Biogenetics Research. Academic Press, USA.
- Harlow E, Lane D (1988): Labelling antibodies with fluorochrome antibodies. A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory.
- Henderson KS, Jackwood DJ (1990): Comparison of the dot blot hybridization assaywith antigen detection assays for the diagnosis of ınfectious bursal disease virus infections. Avian Dis, 34, 744-748.
- Jeffrey IS (1982): Fluorescencemicroscopy. Int Lab, 12, 46-52.
- Kumar A, Rao, AT (1993): Immunofluorescent studies on infectious bursal disease in chickens. Indian Vet J, 15, 26- 29.
- Lukert PD, Saif YM (1991): Infectious bursal disease. 648-668. In: Disease of Poultry. Calnek BW (Ed). 9th ed. Iowa State University Press. Ames, Iowa. USA.
- Maestrone G, Coffin DL (1964): Study of Newcastle disease by means of fluorescent antibody technique. Am J Vet Res, 25, 217-223.
- Pala HH, Türe O (1996): Gumboro hastalığının immunperoksidaz ve immunfloresan teknikleri ile teşhisi. Bornova Vet Kont Araş Enst Md Derg, 21, 161-175.
- Rosales GA, Villegas P, Lukert D, Fletcher JO, Brown J (1989): Immunosupressive potential and pathogenecity of a recent isolate of infectious bursal disease virus in commercial broiler chickens. Avian Dis, 33, 724-728.
- Van der Berg TP, Eterradossi N, Toquin D, Meulemans G. (2000): Infectious bursal disease (Gumboro disease). Rev Sci Tech Off Int Epiz, 19, 527-543.
Öz
Bu çalışmada, infeksiyöz bursal hastalığının (IBD) teşhisinde immunofloresan tekniğinin kullanılması, standardizasyonu ve bu teknikte kullanılacak konjugatın hazırlanması amaçlandı. İmmunize edilen tavuk ve tavşan serumları floresein izotiosiyanat ile işaretlendi. Konjugat standardizasyonu için deneysel infekte ve kontrol grubundan hazırlanan preparatlarla yapılan testlerde optimal konjugat sulandırmasının 1:16 olduğu belirlendi. Ayrıca standardizasyon çalışmaları sonrasında indirekt floresan antikor tekniğinin (IFAT) relatif olarak direkt floresan antikor tekniğine (FAT) üstünlüğü saptandı. AGID testinde kontrol grubuna ait örnekler negatif, aşılı ve saha suşu ile infekte edilen gruplara ait örnekler ise pozitif sonuç verdi. AGID’e göre FAT ve IFAT’ın spesifite ve sensitivitesinin %100 olduğu hesaplandı. Saha çalışmalarında, IBD şüpheli 27 kümesin 23’ünde (%85.13) pozitiflik saptandı. Sonuçta, immunofloresan tekniğinin IBDV infeksiyonlarında kullanılabilirliği ortaya konuldu
Anahtar Kelimeler
Kaynakça
- Akan M (2002): İnfeksiyöz bursal hastalık. 169-178. In: Kanatlı Hayvan Hastalıkları. İzgür M, Akan M. (Eds), Medisan Yayınevi, Ankara.
- Allen, GM, McNulty MS, Connor TJ, McCracken RM, McFerran JB (1984): Rapid diagnosis of infectious bursal disease infection by immunoflourescence on clinical material. Avian Pathol, 13, 419-427.
- Anonim (1996): Infectious bursal disease (Gumboro disease). Chapter 3. 6. 1. In: Manual of Standarts for Diagnostic Tests and Vaccines. OIE Lists A and B diseases of mammals, birds and bees. 3th edition, Paris. An immunoperoxidase monoclonal antibody stain for rapid diagnosis of infectious bursal disease. Avian Dis, 31, 538- 545.
- Cruz-Coy JS, Giambrone JJ, Panangala VS (1993): Production and characterization of monoclonal antibodies against variant A infectious bursal disease virus. Avian Dis, 37, 406-411.
- Goldman M, Carver RK (1957): Preserving flourescein isocyanate for simplified preparation of fluorescent antibody. Science, 126, 839-840.
- Goldman M (1968): Fluorescent Antibody Methods. Biogenetics Research. Academic Press, USA.
- Harlow E, Lane D (1988): Labelling antibodies with fluorochrome antibodies. A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory.
- Henderson KS, Jackwood DJ (1990): Comparison of the dot blot hybridization assaywith antigen detection assays for the diagnosis of ınfectious bursal disease virus infections. Avian Dis, 34, 744-748.
- Jeffrey IS (1982): Fluorescencemicroscopy. Int Lab, 12, 46-52.
- Kumar A, Rao, AT (1993): Immunofluorescent studies on infectious bursal disease in chickens. Indian Vet J, 15, 26- 29.
- Lukert PD, Saif YM (1991): Infectious bursal disease. 648-668. In: Disease of Poultry. Calnek BW (Ed). 9th ed. Iowa State University Press. Ames, Iowa. USA.
- Maestrone G, Coffin DL (1964): Study of Newcastle disease by means of fluorescent antibody technique. Am J Vet Res, 25, 217-223.
- Pala HH, Türe O (1996): Gumboro hastalığının immunperoksidaz ve immunfloresan teknikleri ile teşhisi. Bornova Vet Kont Araş Enst Md Derg, 21, 161-175.
- Rosales GA, Villegas P, Lukert D, Fletcher JO, Brown J (1989): Immunosupressive potential and pathogenecity of a recent isolate of infectious bursal disease virus in commercial broiler chickens. Avian Dis, 33, 724-728.
- Van der Berg TP, Eterradossi N, Toquin D, Meulemans G. (2000): Infectious bursal disease (Gumboro disease). Rev Sci Tech Off Int Epiz, 19, 527-543.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Veteriner Cerrahi |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 1 Eylül 2007 |
| Yayımlandığı Sayı | Yıl 2007Cilt: 54 Sayı: 3 |