PPRV enfeksiyonunun epidemiyolojisinin N ve F genlerine dayalı farklı PCR teknikleri ile araştırılması

Bu calismada, Turkiye’nin degisik bolgelerinde yetistirilen 6 ay yasin uzerindeki koyun ve kecilerden PPRV enfeksiyonu suphesi ile alinan kan, nasal swap ornekleri ile Pendik Veteriner Kontrol Enstitusu Viral Teshis Laboratuvarina doku ornekleri (lenf yumrusu, akciger, rektum, ince barsak ve dalak) RT-PCR, kan serum ornekleri Competative ELISA (C-ELISA) teknikleri ile kontrol edilmistir. Asili ve asisiz olarak 2 grup hayvandan alinan kan serum orneklerinin kontrolu sonucunda, asili hayvanlarda seropozitiflik koyunlarda %12,8, kecilerde %44, asisiz hayvanlarda ise %90,3 olarak bulunmustur.Virolojik kontrol amaciyla alinan toplam 574 ornekten viral nukleik asit tespiti F ve N gen bolgelerini hedef alan konvansiyonel RT-PCR ve N gen bolgesi hedef alan Real-Time RT-PCR ile yapilmistir. Konvansiyonel F gen RT-PCR ile %35,5 (204/574), konvansiyonel N gen RT-PCR ile %39,3 (226/574) ve Real Time N gen RT-PCR ile %44,4 (255/574) oraninda pozitiflik elde edilmistir.  Uc metot birlikte degerlendirildiginde kontrol edilen orneklerin %45.8’i (263/574) pozitif olarak belirlenmistir. Pozitiflik oranlari yillara gore degerlendirildiginde, 2010 yilinda %45,9; 2011 yilinda %45,7 ve 2012 yilinda %45,9 olarak dagilim gostermektedir.Konvansiyonel RT-PCR testleri sonunda elde edilen pozitif orneklerden illeri temsil edecek sekilde, 53 adet F gen ve 60 adet N gen kismi dizin analizleri yapilmis ve gen bankasi ulasim numaralari JQ388615-JQ388664, JQ519907-JQ519965,  JX117877-JX117880 olarak 3 grup altinda alinmistir. Yapilan filogenetik analiz sonrasinda, her iki gen bolgesine gore ulkede sirkule olan PPR viruslarinin IV. genetik hatta yer aldigi gorulmustur.  F gen kismi dizin analizleri sonunda elde edilen dizinlerin nukleotid duzeyinde kendi aralarinda, Turkey2000 ve Nigeria/75/1 dizini ile sirasiyla %98,2-100, %97,9-98,9 ve %91,3-92,4 oraninda benzer olduklari tespit edilmistir. N gen kismi dizin analizleri sonunda ise dizinlerin nukleotid duzeyinde kendi aralarinda, Turkey2000 ve Nigeria/75/1 dizini ile sirasiyla %94,2-100, %94,2-98,3 ve %89,3-90,9 oraninda benzer olduklari tespit edilmistir.   Bu sonuclar isiginda PPRV enfeksiyonunun ulkede endemik oldugu bir kez daha ortaya konmus; enfeksiyonun tanisinda rRT-PCR tekniginin, kullanilan konvansiyonel tekniklere gore etkinligi gosterilmis, hayvan hareketlerinin sinirlandirilmasinin zorluguna dikkat cekilerek enfeksiyondan korunmada en etkili yolun duyarli populasyonlarin asilanmasi oldugu sonucuna varilmistir.AbstractIn this study, from sheep and goats which was grown in different regions of Turkey over the age of 6 months blood, nasal swaps was taken for the potential of PPRV infection occurence. Also  tissue and serum samples ((lymph node, lung, rectum, small intestine, and spleen)  sent to Pendik Veterinary Control Institute, Viral Diagnostic Laboratory was  checked with RT-PCR and Competative ELISA (C-ELISA) techniques respectively. As a result of control of blood serum samples from two groups of animals vaccinated and unvaccinated, seropositivity was found as 12.8% in vaccinated sheep, 44% in vaccinated goats and was found to be 90.3% in unvaccinated animals.            Nucleic acid detection was made with F and N gene targeting conventional RT-PCR and N gene targeting real-time RT-PCR on 574 samples taken for virological control. The conventional RT-PCRs which was made for F and N the positivity found as 35.5% (204/574), 39.3% (226/574) respectively and 44.4% (255/574 ) with N gene real time RT-PCR.  When the three methods considered together the positivity rate of  samples were found as 45.8% (263/574). The review of the positivity rates according to years is 45.9% in 2010, 45.7% in 2011 and 45.9% in 2012 .         From the positive samples obtained from conventional RT-PCRs, 53 F and 60 N genes partial sequence analysis that representing all the provinces was made and taken under the three groups according to the gene bank accession numbers (JQ388615-JQ388664, JQ519907-JQ519965, JX117877-JX117880). After the phylogenetic analysis, it was seen that PPR viruses circulated in the country take place  in the IV. genetic line according to the both genes. At the end of the F gene partial sequence analysis, it was found 98.2 to 100%, 97.9 to 98.9% and 91.3 to 92.4% similar with each other, Turkey2000 and Nigeria/75/1 sequences respectively. At the end of the N gene partial sequence analysis, it has been identified %94,2-100, %94,2-98,3 ve %89,3-90,9 similar with each other Turkey2000 and Nigeria/75/1 sequences respectively.    In the light of these results it has been revealed once again that PPRV infection is endemic in the country and in the diagnosis of infection, efficiency of  rRT-PCR technique was shown compared to the conventional techniques. As a result it was concluded that vaccination of susceptible populations is the the most effective way to the prevention of infection because of the difficulty in restriction of animal movements.