Farklı PCR Sistemlerinin Tavuk Trakeasından Mycoplasma gallisepticum Tespiti için Değerlendirilmesi

Bu çalışmada, Mycoplasma gallisepticum (MG) ile doğal enfekte damızlık tavuklarda kümeslerin MG seropozitivitesine çabuk serum aglütinasyon testiyle bakıldı, MG-DNA'sının tespitinde; Air Thermal Cycler (ATC) PCR (Idaho Technologies) ve hızlı, güvenilir bir metot olan LightCycler real-time PCR sistemleri (LC PCR) (Roche Diagnostics, Manheim, Germany) kullanıldı ve MG LC PCR'ın hem saf kültür ve hem deneysel olarak kontamine edilmiş örneklerden MG DNA'sını tespit etme limiti belirlendi. Üç farklı firmaya ve 16 kümese ait 177 trakeal swap LC PCR ile çalışıldı. On kümesten 177 tavuk MG-seropozitif olarak teşhis edilmesine rağmen bunlardan sadece 3 (30%) kümesten alınan 41 (%35) tracheal örnek LC PCR ile pozitif bulundu. Altı MG-seronegatif kümesten alınan 60 örnek (%33.8) aynı zamanda LC PCR ile de MG negatif olarak bulundu. Daha önce aynı kümeslerin de dahil olduğu 4 farklı firmadan alınan 212 MG-seropositive örneklerden sadece 4 tanesi (%1.8)'i ATC PCR ile pozitif bulunmuştu. Geliştirdiğimiz LC PCR, DNA ekstraksiyonu ile birlikte yaklaşık olarak 6 saatte sonuç vermektedir. Aynı zamanda kanatlı işletmelerinde MG-enfekte kümeslerin taranmasında kullanılabilecek hızlı ve güvenilir, hazır bir tespit ve doğrulama sistemidir.

Evaluation of Different PCR Systems for the Detection of Mycoplasma gallisepticum in Chicken Trachea

In this work, we detected the MG-serologic condition by rapid plate agglutination tests, used Air Thermal Cycler (ATC PCR) (Idaho Technologies) and LightCycler real-time PCR system (LC PCR) (Roche Diagnostics, Manheim, Germany) for rapid and reliable detection of Mycoplasma gallisepticum (MG) from tracheal swab samples of naturally infected breeder chickens, and determinated MG-DNA detection limit by MG LC PCR from both pure culture and artificially spiked samples. One hundred and seventy seven tracheal swab samples from 16 flocks of 3 different companies were tested by LC PCR. Despite 117 chickens from 10 flocks were diagnosed as MG-seropositive, only 41 (35%) of tracheal swab samples from 3 (%30) flocks were found positive by LC PCR. Sixty (33.8%) of the samples from 6 MG-seronegative flocks were also found to be MG negative by LC PCR. Two hundred twelve MG-seropositive samples from 4 companies (3 of them are same companies tested previously by LC) were tested by ATC PCR and detected only 4 (1.8%) tracheal swab samples MG-positive. The LC PCR gives the results in approximately 6 hours DNA extraction, and is rapid and reliable confirmation and detection test ready to be implemented for screening MG-infected flocks in poultry companies.

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