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

Year 2016, Volume: 27 Issue: 3, 135 - 139, 20.12.2016

Sevil Erdenliğ Gürbilek Oktay Keskin , Esra B Üyükçangaz Osman Yaşar Tel

Abstract

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

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

Abstract

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.

Keywords

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

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