Mycoplasma gallisepticum ve Mycoplasma synoviae için Tanısal Kantitatif Gerçek Zamanlı PCR Geliştirilmesi

Öz

Kanatlı mikoplazma infeksiyonları, solunum yolu hastalığı, sinovitis ve düşük performans ile ilişkili ekonomik problemlere neden olur. Kanatlı mikoplazmalarının teşhisinde temel yaklaşım bakterinin izolasyon ve identifikasyonudur. Ancak kullanılan konvansiyonel bakterinin nazlı üreme şeklinden dolayı zaman alıcı, zor ve tecrübeli personele gereksinim gösteren bir yöntem olması, alternatif teşhis yöntemlerinin önemini ön plana çıkarmıştır. Bu nedenle, bu çalışmada, Mycoplasma gallisepticum (MG) ve Mycoplasma synoviae (MS)’nin teşhisine yönelik kantitatif gerçek zamanlı PCR (qPCR) yöntemi geliştirilmesi amaçlandı. Bu amaçla, M. gallisepticum’un teşhisi için lipoprotein (lp), M. synoviae’nın teşhisi lipoprotein hemaglütinin (vlhA) genleri seçildi. Yapay kontaminasyon yapılan svap örneklerinde, geliştirilen metodun deteksiyon limiti (LOD) 100- 101 DNA/µl olarak belirlendi. Spesifite ve sensitivite oranları ise %100 olarak tespit edildi. Tüm sonuçlar birlikte değerlendirildiğinde, geliştirilen qPCR metodunun MG ve MS’nın hızlı ve doğru teşhisinde kullanılabilecek bir yöntem olduğu belirlendi.

Anahtar Kelimeler

• Mycoplasma gallisepticum
• Mycoplasma synoviae
• qPCR

Development of Diagnostic Quantitative Real-Time PCR for Mycoplasma gallisepticum and Mycoplasma synoviae

Öz

Avian mycoplasmas are associated with respiratory disease, synovitis, poor quality of day-old chicks and poor performance. The main approach used for the diagnosis of avian mycoplasmas is isolation and identification of the microorganism. Since the bacteria are slow growing fastidious organisms, conventional methods are time consuming, laborious and require experienced personnel. For this reason, we aimed to develop a rapid detection method for Mycoplasma gallisepticum and Mycoplasma synoviae by quantitative real-time polymerase chain reaction (qPCR). For this purpose, the lipoprotein (lp) and variable lipoprotein hemagglutinin (vlhA) genes were used to detect M. gallisepticum and M. synoviae, respectively. The limit of detection (LOD) of the assay was determined to be 100- 101 DNA/µl from artificially contaminated swab samples. The specificity and sensitivity ratios were 100%. Overall, these results indicate that this qPCR method can be accurately used for detection of MG and MS.

Anahtar Kelimeler

• Mycoplasma gallisepticum
• Mycoplasma synoviae
• qPCR

Kaynakça

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