OKTAY GENÇ, ÖZLEM BÜYÜKTANIR YAŞ, Nevzat YURDUSEV

Development of qualitative and quantitative ELISA models for bovine Brucellosis diagnosis

Bu çalışmada, IgG izotipi anti-Brucella abortus LPS antikorlarının taranması ve nicelendirilmesi için kalitatif ve kantitatif ELISA modelleri (iELISA ve qELISA) geliştirildi. Yüksek düzey duyarlılık ve özgünlüğe sahip ELISA modellerinin geliştirilmesinde doyurma-dilüsyon reaktiflerinin tabiatı ve konsantrasyonları çok kritik bulunmuştur. Bu çerçevede, süt bileşenlerinden αlaktalbuminin inhibitör etkisi bulunmadığı ve tanımlanmamış bir mekanizmadan dolayı kazein ve özellikle β-laktoglobulinin anti-LPS antikor tespitini güçlü olarak inhibe ettiği belirlenmiştir. iELISA ve qELISA modelleri OIE referans serumlarının yanı sıra RBT, CFT ve kompetitif ELISA ile değerlendirilerek belirlenmiş saha serumları yardımıyla geliştirilerek valide edilmiştir. Elde edilen sonuçlar, bu çalışmada geliştirilen her iki ELISA modelinden iELISA’nın geniş ölçek taramalara ve qELISA’nın ise anti-LPS antikor konsantrasyonunun standart bir şekilde belirlenmesine yönelik olarak, endemik ülkelerde brusellozis taramaeradikasyon ve aşı izleme programlarında etkin ve geçerli immunodiyagnostik araçlar olarak kullanılabileceğini göstermektedir.

Anahtar Kelimeler:

sığır, beta-laktoglobulin, brusellozis, kazein, teşhis, tanı teknikleri, hastalık kontrolü, hastalık önleme, ELISA, IgG, bağışıksal tanı, nitelik analizleri, nicelik analizleri, Brucella abortus

Sığır Brusellozis tanısı için kalitatif ve kantitatif ELISA modellerinin geliştirilmesi

In this study, a qualitative and a quantitative ELISA models (iELISA and qELISA, respectively) have been developed for screening and quantifying IgG isotype anti-Brucella abortus LPS antibody. For developing high sensitive and specific ELISA models, the nature and concentration of blocking-diluting reagents have been found very critical since milk components such as casein and particularly β-lactoglobulin, but not α-lactalbumin, strongly inhibited the anti-LPS antibody detection by an unknown mechanism. The iELISA as well as qELISA models have been developed and validated by OIE reference sera and well-known field sera evaluated with RBT, CFT and cELISA. The results demonstrated that both ELISA models, iELISA in large-scale screening and qELISA in standard determination of anti-LPS antibody concentration, developed in this study can be included as valuable and effective immunodiagnostic tools in brucellosis monitoring-eradication and vaccination surveillance programs in endemic countries.

Keywords:

cattle, beta-lactoglobulin, brucellosis, casein, diagnosis, diagnostic techniques, disease control, disease prevention, ELISA, IgG, immunodiagnosis, qualitative analysis, quantitative analysis, Brucella abortus,

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