Development of an In House Enzyme-Linked Immunosorbent Assay (ELISA) Prototype Using Different Anti- gens for Serological Diagnosis of Brucella ovis

Abstract

In this study, it was aimed to develop an Enzyme-Linked Immunosorbent Assay (ELISA) prototype to evalua- te humoral immune response against Brucella ovis by using different antigenic fractions from bacteria comparatively. For this purpose, four different antigens, namely Hot saline extract, Rough Lipopolysaccharide, Phage lysate 1, and Phage lysate 2 were used to test 84 sheep serum samples in ELISA. Seropositivity rates were, 14.3%, 13.1%, 10.7%, and 15.5%, respectively. Although the highest seropositivity rate was obtained by using the Phage lysate 2 antigen, it did not reach the level of statistical significance (χ2=0.89; P>0.05) difference between these antigens. It was concluded that in future studies, there would be a need for comparing the results of more serum samples tested in ELISA using Phage lysate 2 with the results of standard serological tests such as complement fixation test and agar gel immunodif- fusion test or available commercial ELISA kit. Thus, the potential possibility of commercialization of a new in-house ELISA prototype will be evaluated more realistically. Given the seropositivity rate of Brucella ovis infection obtained by using a small number of serum samples, a large-scale serological study with greater serum samples from different parts of Turkey will reveal the real disease situation in our country.

Keywords

• sheep

Brucella ovis’in Serolojik Tanısı Amacıyla Farklı Antijenlerin Kullanıldığı Bir In House Enzyme- Linked Immunosorbent Assay (ELISA) Prototipinin Geliştirilmesi

Abstract

Bu çalışmada, Brucella ovis’e karşı humoral yanıtın değerlendirilmesinde kullanılmak üzere, bu bakterinin farklı antijenik fraksiyonlarından hazırlanan antijenlerin karşılaştırılmasıyla bir Enzyme -Linked Immunosorbent Assay (ELISA) prototipinin geliştirilmesi amaçlandı. Bu amaçla, dört farklı antijen (Hot salin ekstrakt, Rough Lipopolisakkarit, Faj lizat 1 ve Faj lizat 2), 84 koyun serum örneği ile test edildi. Brucella ovis açısından ELISA seropozitiflikleri, kullanı- lan antijenlere göre sırasıyla %14.3, %13.1, %10.7 ve %15.5 olarak saptandı. En yüksek pozitiflik oranı Faj lizat 2 ile elde edildi, ancak, Brucella ovis’e karşı oluşan antikorların ELISA ile belirlenmesi için kullanılan 4 farklı antijenin test sonuçları arasındaki farklılık, istatistiki açıdan önemsiz (χ2=0.89; P>0.05) bulundu. Sonuç olarak, gelecekteki çalışma- larda, Faj lizat 2 antijeni kullanılarak ELISA ile test edilen çok daha fazla serum örneği için elde edilen sonuçların, komplement fikzasyon testi ve agar jel immunodifüzyon testi gibi standart testler ve temin edilebilecek ticari kitlerin sonuçları ile karşılaştırılmasına ihtiyaç duyulacağı ve böylece in house ELISA prototipinin ticarileşme potansiyelinin daha sağlıklı olarak değerlendirilebileceği kanısına varıldı. Ayrıca, az sayıda serum örneğiyle elde edilen seropozitiflik oranı göz önünde bulundurulduğunda, Türkiye'nin farklı bölgelerini kapsayan daha fazla sayıda serum örneği ile geniş ölçekli bir serolojik çalışmanın yapılması ile ülkemizdeki gerçek hastalık durumunun ortaya çıkarılabileceği düşünül- mektedir.

Keywords

• Brucella ovis
• ELISA
• koyun
• sheep

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