SERPİL KAHYA DEMİRBİLEK, Özge YILMAZ, Ayşegül EYİGÖR, SERAN TEMELLİ, K. Tayfun ÇARLI

Detection of Mycoplasma gallisepticum and Mycoplasma synoviae by Real-Time PCRs and Mycoplasma gallisepticum-antibody Detection by an ELISA in Chicken Breeder Flocks

Bu çalışmada, Mycoplasma gallisepticum (MG) ve Mycoplasma synoviae (MS)'nın solunum sistemine ait semptomlara sahip damızlık tavuk kümeslerindeki yaygınlığını tespit etmek amaçlandı. Toplam 77 kümes (2153 trakeal svab ve kan örneği)'in toplanan tüm örnekleri MG gerçek zamanlı PCR (MG-rPCR) ve MG-ELISA ile, 32 kümes ise MS-rPCR ile test edildi. Çalışmanın birinci bölümünü kapsayan ilk 28 kümeste MG-antikorları yüksek bulundu ve belirgin klinik semptomlar gösteren bu tavukların örnekleri MG-rPCR1 ile negatif sonuç verdi. Bu nedenle, bu PCR'da kullanılan MG-lipoprotein geni-spesifik primerler (MG-rPCR1), MS tespitinde kullanılan MS-16S rRNA primerlerine (MS-rPCR) benzer olarak, MG-16S rRNA primerleri (MG-rPCR2) ile değiştirildi ve çalışmaya bu primerler ile devam edildi. MG-ELISA ile %100 pozitif olan ilk 28 kümesin tümü MG-rPCR1 ile negatif iken, çalışmanın 2. bölümünde %87.8 MG seropozitif olarak saptanan diğer 49 kümes MG-rPCR2ile %42.9 pozitif bulundu. Ayrıca, bu ilk 28 kümesten seçilen 5 adetinde MS-rPCR negatif iken, daha sonraki 49 kümesten seçilen 27'si MSrPCR ile %7.4 pozitif olarak tespit edildi. Genel olarak, toplam 77 kümeste uygulanan 432 MG-rPCR1-2'den 81'i (%18.7) ve bu kümeslerden 32 adetinde yapılan 187 MS-rPCR'ın 13'ü (%6.9) pozitif bulundu. ELISA sonuçları ise serolojik testlerden önemli oranda yanlış-pozitif sonuçlar alınabileceğini ve tek bir test sistemine güvenilmemesi gerektiğini işaret etti. Bu çalışma ile, aynı zamanda, Mycoplasma-enfekte kümeslerin laboratuvarlarda doğrulanabilmesi için doğru primerler seçildiğinde rPCR'ın güvenli bir metot olduğu, damızlık tavuk kümeslerinde MG ve MS prevalansının özellikle kış mevsiminde oldukça yüksek olduğu ortaya kondu

Damızlık Tavuk Kümeslerinde Mycoplasma gallisepticumve Mycoplasma synoviae'nin Gerçek Zamanlı PCR'lar ile ve Mycoplasma gallisepticum Antikorlarının ELISA ile Tespiti

This study aimed to determine the prevalence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) in breeder flocks showing respiratory symptoms. A total of 77 flocks (2153 tracheal swabs and blood samples) were sampled and all were tested by MG real time PCR (MG-rPCR) and MG-ELISA, and 32 flocks were tested by MS real time PCR (MS-rPCR). In the first part of this study covering 28 flocks, all samples from chickens with marked clinical symptoms and high MG-antibody levels gave negative results with MG-rPCR1. Therefore, the MG-lipoprotein gene-specific primers (MG-rPCR) of this PCR were replaced with MG-16S rRNA primers (MG-rPCR2), as were the MS16S rRNA primers (MS-rPCR), thus the study was pursued accordingly. All of the first 28 flocks, which were 100% positive by MG-ELISA, were MG-rPCR1 negative, whereas in the second part of the study, other 49 flocks, which were 87.8% MG seropositive, were found 42.9% positive by MG-rPCR2. In addition, 5 selected flocks from the first 28 were negative, whereas 7.4% of the 27 selected flocks from the second 49 were positive by MS-rPCR. Overall, 81 out of 432 MG-rPCR1-2 (18.7%) performed from 77 flocks, and 13 out of 187 MS-rPCRs (6.9%) in 32 flocks were determined as positive. ELISA results indicated that there could be significantly high false-positives in serological tests, thus results should not be relied upon one test system. Also, this study revealed that, for the confirmation of Mycoplasma-infected flocks in laboratories, rPCR is a reliable method as long as suitable primers are selected, and that MG and MS prevalence is considerably high in winter season

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