Sevil ERDENLİG, Ümit SEVİMLİ, Osman Yaşar TEL

Solunum Bozukluğu olan Sığırlarda Bulaşıcı Sığır Pleuropnömonisine Karşı Oluşan Hümoral İmmün Yanıtın Western Blot Tekniği, Kompleman fiksasyon testi ve Kompetatif Enzyme Linked İmmunosorbent Assay Testi ile Değerlendirilmesi

Bu çalışmada, solunum bozuklukları gösteren sığırlardan alınan 253 serum numunesi, doğrulayıcı olarak Western Immunoblotting test (IBT), Kompleman fiksasyon testi (CFT) ve kompetatif enzyme linked immunosorbent assay (c-ELISA) ile test edildi. CFT ile 253 serum örneğinden ikisi (%0,8) pozitif, ikisi (%0,8) şüpheli, c-ELISA ile 7 (%2,8) serum numunesi pozitif ve 25'i (%9,9) şüpheli bulundu. Öte yandan, 110, 98, 95, 62/60 ve 48 kDa' olarak tanımlanması gereken antijenik bantlar, serum örneklerinin hiçbirinde tespit edilmedi. Bazı serum numunelerinin İmmünoblot analizi sonucunda bir veya üç arasında bant görüldü ve bu bantların bazıları oldukça soluktu. MmmSc suşları arasındaki elektroforetik profiller ile ilgili benzerlikleri tespit etmek için sığırlardan izole edilen Mycoplasma türlerinin tam hücre proteinleri ve MmmSC referans suşları (Mycoplasma mycoides subsp. mycoides SC Botswana (Afrika suşu), Mycoplasma mycoides subsp. mycoides SC PG1 (referans suş), Mycoplasma mycoides subsp. mycoides SC V5 (Australian strain), M. bovigenitalium, M. bovirhinis, M. capricolum subsp. capricolum, M.canadense, M.alkalescens and M. bovis). sodium dodecyl sulphate polyacrilamide gel elektroforezi (SDS-PAGE). ile analiz edildi The result of this analysis indicated that antigens of 220, 78, 82, 68, 24 kDa were common among all tested strains. According to these results, it was thought that positive results by both CFT and c-ELISA, also non specific band patterns observed by IBT might be caused by a cross reaction between tested mycoplasma strains. As conclusion, it is recommended that all the positive results by CFT and c-ELISA be confirmed by IBT. Bu analizin sonucu, 220, 78, 82, 68, 24 kDa antijenlerinin test edilen tüm suşlar arasında yaygın olduğunu gösterdi. Bu sonuçlara göre, CFT ve c-ELISA pozitif sonuçlarının ve IBT ile gözlemlenen spesifik olmayan bant örneklerinin, test edilen mikoplazma suşları arasındaki çapraz reaksiyonlardan kaynaklanabileceği düşünüldü., CFT ve c-ELISA ile elde edilen tüm pozitif sonuçların IBT testi ile doğrulanması gerektiği sonucuna varıldı.

Anahtar Kelimeler:

ELISA, Immunoblotting test, Seroloji

The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test

In this study 253 serum samples from cattle showing respiratory disorders were tested by confirmatory Western Immunoblotting test (IBT), complement fixation test (CFT) and competitive enzyme linked immunosorbent assay (c-ELISA). Two (0.8%) out of 253 serum samples were found to be positive and two were (0.8%) doubtful by CFT while 7 (2.8%) serum samples were found as positive and 25 (9.9%) of them were found to be doubtful by c-ELISA. On the other hand, a core profile of antigenic bands needed to be identified at 110, 98, 95, 62/60 and 48 kDa was not detected in any of the test serum sample. Immunoblot analysis some of serum samples displayed the number of bands between one or three and some of them were highly faint. In order to detect similarities between MmmSc strains related to their electrophoretic profiles, whole cell proteins of Mycoplasma species isolated from cattle and MmmSC reference strains (Mycoplasma mycoides subsp. mycoides SC Botswana (African strain), Mycoplasma mycoides subsp. mycoides SC PG1 (reference strain), Mycoplasma mycoides subsp. mycoides SC V5 (Australian strain), M. bovigenitalium, M. bovirhinis, M. capricolum subsp. capricolum, M.canadense, M.alkalescens and M. bovis) were analysed by sodium dodecyl sulphate polyacrilamide gel elektrophoresis (SDS-PAGE). The result of this analysis indicated that antigens of 220, 78, 82, 68, 24 kDa were common among all tested strains. According to these results, it was thought that positive results by both CFT and c-ELISA, also non specific band patterns observed by IBT might be caused by a cross reaction between tested mycoplasma strains. As conclusion, it is recommended that all the positive results by CFT and c-ELISA be confirmed by IBT.

Keywords:

ELISA, Immunoblotting test, Serology,

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