Sevil ERDENLİĞ GÜRBİLEK, Oktay KESKİN, Esra B ÜYÜKÇANGAZ, Osman Yaşar TEL

Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması

Bu çalışmada, B. canis infeksiyonunun serolojik teşhisi için B. canis M- ve B.abortus RB51 suşlarından hazırlanan antijenlerin kullanıldığı indirekt ELISA (i-ELISA) geliştirilmesi amaçlanmıştır. B. canis infeksiyonu etken izolasyonu ve agar jel immunodiffuzyon (AGID) testi ile doğrulanmış 16 adet pozitif serum ve B. canis infeksiyonu yönünden hiçbir klinik belirti göstermeyen AGID negatif 24 adet serum testin duyarlılığı ve özgüllüğünü saptamak için referans serumlar olarak kullanılmıştır. Validasyon paneli kullanılarak, tanısal duyarlılık ve özgüllüğü %100 verecek eşik değeri B. abortus RB51 i-ELISA için OD450 0.243 ve B. canis M- i-ELISA için OD450 0.354 olarak belirlenmiştir. Saptanan eşik değer göz önüne alınarak, Şanlıurfa ve Bursa Büyükşehir Belediyesi Köpek Barınaklarından temin edilen 113 adet köpek serumu çabuk lam aglütinasyon testi (ÇLAT) ve geliştirilen iki i-ELISA prototipi ile test edilmiştir. Şanlıurfa ilinden temin edilen 47 köpek serumundan 6’sı (%12.8) ÇLAT ile, 3’ü (%6.4) B. abortus RB51 i-ELISA ile ve ikisi (%4.2) de B. canis M- i-ELISA ile pozitif bulunmuştur. Bursa ilinden temin edilen 66 adet köpek serumundan 11’i (%16.6) ÇLAT ile, 5’i (%7.6) B. abortus RB51 i-ELISA ile ve 4’ü (%6.1) de B. canis M- i-ELISA ile pozitif bulunmuştur. Sonuçta B. abortus RB51 i-ELISA ve B. canis M- i-ELISA’nın küçük farklılıklar ile benzer duyarlılık ve özgüllük gösterdikleri ve bu testlerin köpeklerde B. canis infeksiyonunda doğrulayıcı test olarak ÇLAT ile birlikte başarılı bir şekilde kullanılabileceği kanısına varılmıştır.

Anahtar Kelimeler:

Brucella canis M -, Brucella abortus RB51, i-ELISA, seroloji, köpek

Evaluation of Usage Indirect ELISA Using Antigens From Two Different Brucella Strains in Serological Diagnosis of B. canis Infection

The aim of this study was to develop two indirect ELISAs (i-ELISA) using heat-extracted antigens prepared with the B. canis M- and B. abortus RB51 strains for the serologic diagnosis of B. canis infection in dogs. Reference sera consisted of 16 agar gel immunodiffusion (AGID)-positive sera from animals with clinical signs and positive bacterial isolation and 24 AGID-negative sera from healthy animals with no history or evidence of canine brucellosis were used to assess diagnostic sensitivity and specificity of the tests. Using validation panel, the cut-off values of OD450 0.243 and 0.354 were selected to give 100 % diagnostic specificity and sensitivity for B. abortus RB51 i-ELISA and B. canis M- i-ELISA, respectively. Based on these cut-off values, 113 serum samples taken from dog shelters in Şanlıurfa and Bursa metropolitan municipalities tested by rapid slide agglutination test (RSAT) and both i-ELISAs. Of the 47 dog sera tested from Şanlıurfa, 6 (12.8%) were positive by RSAT, 3 (6.4%) were positive by B. abortus RB51-i-ELISA and two of the samples (4.2%) were found positive by B. canis M- i-ELISA. Of the 66 serum samples from Bursa, 11 (16.6%) were found positive by RSAT, 5 (7.6%) and 4 (6.1%) of which were confirmed by B. abortus RB51 i-ELISA and B. canis M- i-ELISA, respectively. In conclusion, both B. abortus RB51 i-ELISA and B. canis M- i-ELISA yielded the same diagnostic specificity and sensitivity with minor differences and it was thought that they could be used successfully as a confirmatory test together with RSAT as a screening test.

Keywords:

Brucella canis M-, Brucella abortus RB51, i-ELISA, Serology, Dog,

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