Ren-Feng LI, Xiang-Qin TIAN, Zi-Liang WANG, Jin-You MA, Li-Rong WANG, Shou-Ping ZHANG, Yan-Yan YANG, Jin-Qing JIANG, San-Hu WANG, Yan YU

Kolostrumda Domuz Epidemik Diare Virus-Spesifik IgA Antikorlarının Belirlenmesinde ELISA Sisteminin Geliştirilmesi ve Değerlendirilmesi

Domuz Epidemik Diare Virus (DEDV) enfeksiyonu domuzların oldukça yıkıcı enfeksiyöz bir hastalığıdır. Virusa karşı bağışıklık durumunu değerlendirmek ve tanıyı koyabilmek için yüksek performanslı metotların gelişimi hastalığın kontrol altına alınabilmesi için elzemdir. Bu çalışmada, kolostrumdaki IgA antikorlarını taramak amacıyla rekombinant S1 protein temelli indirekt enzime bağlı immunosorbent metot (rpS1-iELISA) geliştirildi. rpS1-iELISA için optimize edilmiş eşik değeri 0.448 olarak belirlendi. Sonuçlar %96.87 duyarlılık ve %100 özgüllük gösterdi. Tekrarlanabilirlik testleri kolostrum örnekleri ve koşumlar arasında varyasyon katsayısıının %8’den daha az olduğunu gösterdi. Toplam 523 adet saha kolostrum örneklerinin test sonuçları rpS1-iELISA’nın immunofloresans testi (kappa = 0.958) ile mükemmel uyumlulukta olduğunu ve ticari ELISA kitinden daha iyi performansa sahip olduğunu gösterdi. Mevcut test kolostrumda IgA seviyesini ölçerek DEDV’e karşı mukozal bağışıklık yanıtın koruyuculuk seviyesini tahmin etmede ve tanıyı koymada yardımcı olacaktır.

Development and Evaluation of an ELISA System for Detection of PEDV-Specific IgA Antibodies in Colostrum

Porcine epidemic diarrhea virus (PEDV) is a devastating swine infectious disease. Development of high-performance methods to diagnoseand evaluate viral immune status remains very important to control PEDV. Here, a recombinant S1 protein-based indirect enzyme-linkedimmunosorbent assay (rpS1-iELISA) was developed to monitor IgA antibody in the colostrum. Optimized cut-off value of the rpS1-iELISA wasdetermined as 0.448. Results yielded a sensitivity of 96.87% and a specificity of 100.0%. Repeatability tests indicated that the coefficients ofvariation of the colostrum samples within and between runs were both less than 8%. Test results of 523 field colostrum samples showed thatthe rpS1-iELISA had excellent agreement with immunofluorescence assay (kappa = 0.958) and better test performance than a commercialELISA kit. This test will aid in future diagnostics and assessment of the protective levels of mucosal immune response against PEDV bymeasuring IgA levels in the colostrum.

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