Sığır ve koyun brusellozisinin serolojik teşhisinde konvansiyonel testler ile Kompetitif-ELISA’nın karşılaştırılması

Yıl 2010, Cilt: 21 Sayı: 2, 63 - 70, 30.12.2010

Asiye Dakman Uğur Küçükayan Ufuk Ülker H. Kaan Müştak

Öz

Bu çalışmada Brucella S19 genç aşısı ile aşılanan danalar ile Brucella spp. ile infekte sığırlar ve Brucella Rev1 genç aşısı ile aşılanmış kuzular ile Brucella spp. ile infekte koyunlara ait kan serumları; konvansiyonel testler ve Kompetatif-ELISA (C-ELISA) ile incelenerek, oluşan humoral immun yanıt değerlendirilmiştir. Doğal infekte sığırların %94’ü Rose Bengal Plate Test (RBPT), %98’i Serum Aglütinasyon Test (SAT), %92’si Komplement Fikzasyon Test (CFT) ve %95’i C-ELISA ile pozitif bulunmuştur. Aşı yapılan danalara ait antikor titrelerinde test zamanlarına göre testler arasında farklılıklar meydana gelmiştir. Doğal infekte koyunların %91.16’sı RBPT, %91.16’sı SAT, %85.63’ü CFT ve %86.74’ü C-ELISA ile pozitif bulunmuştur. Sonuç olarak brusellozisin teşhisinde, C–ELISA’nın özellikle CFT’ye alternatif olarak kullanılabileceği gösterilmiştir.

Anahtar Kelimeler

H. Kaan MÜŞTAK, Ufuk ÜLKER, Uğur KÜÇÜKAYAN, Asiye DAKMAN, Brucella, Kompetatif-ELISA, koyun, sığır., 2010, Etlik Veteriner Mikrobiyoloji Dergisi

Comparison of conventional tests with Competitive-ELISA in serological diagnosis of bovine and ovine brucellosis

Öz

In this study, blood sera from calves vaccinated with Brucella S19 vaccine and cattle infected by Brucella spp. and lambs vaccinated with Brucella Rev1 vaccine and sheep infected by Brucella spp. were tested with conventional tests and Competitive-ELISA (C-ELISA) in order to evaluate the humoral immune response. In naturally infected cattle group, 94% with Rose Bengal Plate Test (RBPT), 98% with Serum Agglutination Test (SAT), 92% with Complement Fixation Test (CFT) and 95% with C-ELISA were found positive. Differences between the tests were found in antibody titers on the basis of testing times. In naturally infected sheep group, 91.16% with RBPT, 91.16% with SAT, 85.63% with CFT and 86.74% with C-ELISA were found positive. It was concluded that, in the diagnosis of brucellosis C-ELISA could be used as an alternative test especially to CFT.

Anahtar Kelimeler

Brucella, cattle, Competitive-ELISA, sheep.

Kaynakça

• Agresti A (2002). Categorical data analysis. 2nd edition. Wiley-Interscience Publishers, New York.
• Alton GG, Jones LM, Pietz DE (1975). Laboratory Techniques in Brucellosis. 2nd ed. Geneva, World Health Organisation, Monograph Series, No: 55.
• Alton GG, Jones LM, Angus RD, Verger JM (1988). Techniques for the Brucellosis laboratory. I.N.R.A. Paris.
• Anon (1990): Brucellosis mücadele talimatnamesi. III.Baskı, Hayv. Araşt. Ens.Md.lüğü Ofset Tesisleri., Lalahan – Ankara, No:189.
• Anon (2009a). Bovine brucellosis. In: OIE Manual of diagnostic tests and vaccines for terrestrial animals. Vol. 2, section 2.4, chapter 2.4.3.
• Anon (2009b). Caprine and ovine brucellosis (excluding Brucella ovis). In: OIE Manual of diagnostic tests and vaccines for terrestrial animals. Vol. 2, section 2.7, chapter 2.7.2.
• Diaz-Aparicio E, Aragon V, Marin C, Alonso B, Front M, Moreno E, Perez-Ortiz S, Blasco JM, Diaz R, Moriyon I (1993). Comperative analysis of brucella serotype A and M and Yersinia enterocolitica 0:9 polysaccharides for serological diagnosis of brucellosis in cattle, sheep and goats. J Clin Microbiol, 31(29): 3136-3147.
• Fekete A, Bantle JA, Halling MS, Sanborn MR (1990). Preliminary development of a diagnostic test for brucella using polymerase chain reaction. J App Bacteriol, 69: 216-227.
• Gall D, Colling A, Marino O, Moreno E, Nielsen K, Perez B, Samartino L (1998). Enzyme immunoassays for serological diagnosis of bovine brucellosis: A trial in Latin America. Clin Diag Lab Immunol, 5(5): 654-661.
• Mac Millan A (1990). Conventional serological tests. In: Animal Brucellosis. Eds: Nielsen K, Duncan JR, CRC press Boca Raton Florida. 154-197.
• Marin CM, Moreno E, Moriyon I, Diaz R, Blasco JM (1999). Performance of competitive and indirect enzyme-linked immunosorbent assays, gel immunoprecipitation with native hapten polysaccharide, and standard serological tests in diagnosis of sheep brucellosis. Clin Diagn Lab Immunol, 6, 269-272.
• Nielsen K. (2002). Diagnosis of brucellosis by serology. Vet Microbiol 90: 447-459.
• Nielsen KH, Kelly L, Gall D, Nicoletti P, Kelly W (1995). Improved competitive enzyme immunoassay for diagnosis of bovine brucellosis. Vet Immunol Immunopathol, 46(3-4): 285-291.
• Quinn PJ, Carter ME, Markey B, Carter GR (1994). Clinical Veterinary Microbiology. Wolfe publishing, Spain.:261-267.
• Samartino L, Gall D, Gregoret R, Nielsen K (1999). Validation of enzyme-linked immunosorbent assays for the diagnosis of bovine brucellosis. Vet Microbiol, 70 (3-4): 193-200.
• Stack JA, Perrett LL, Brew SD (1999). Competitive ELISA for bovine brucellosis suitable for testing poor quality samples. Vet Rec, 145: 735-736.
• Stournara A, Minas A, Bourtzi-Chatzopoulou E, Stack J, Koptopoulos G, Petridou E, Sarris K (2007). Assessment of serological response of young and adult sheep to conjunctival vaccination with Rev-1 vaccine by fluorescence polarization assay (FPA) and other serological tests for B. melitensis. Vet Microbiol, 119(1): 53-64