Faculty of Veterinary Medicine University of Zagreb
Abstract
A total of 34 clinical samples were collected for detection of Marek’s disease virus (MDV) by polymerase chain reaction assays using primers M1.1/M1.8 to amplify a region of the ICP4 gene in layer birds of poultry. Primer set M1.1/M1.8 amplified a 318 bp product as against the expected 247 bp product in 30 samples out of 34 samples tested. To confirm the result, this primer was subjected to NCBI BLAST, and it was found that the primer specific segment of 318 bp does exist in the published sequence of Md5 and Md11BAC. The PCR product was sequenced and resulted in 273bp by direct sequencing. The sequence was analysed using NCBI blast and Clustal W with the published sequence of Gallid herpes virus-2 giving a matching score of 97, 96 and 90% indicating a highly conserved region. This shows that the MDV is prevalent in Gujarat.Prikupljena su 34 uzorka kliničkoga materijala nesilica za dokaz virusa Marekove bolesti lančanom reakcijom polimerazom uporabom početnica M1.1/M1.8 za umnažanje područja gena ICP4. Uporabom seta početnica M1.1/M1.8 umnožen je proizvod od 318 bp u odnosu na očekivani od 247 bp u 30 od 34 pretražena uzorka. Za potvrdu rezultata početnica je bila analizirana pomoću programa NCBI BLAST, te je ustanovljeno da početnica za specifičan odsječak od 318 bp postoji u objavljenoj sekvenci Md5 i Md11BAC. Proizvod PCR-a
bio je izravno sekvenciran te je ustanovljeno da sadrži 273 bp. Slijed je bio analiziran uporabom programa NCBI BLAST i Clustal W i uspoređen s objavljenim slijedom za kokošji herpesvirus 2 te je ustanovljena podudarnost od 97, 96 i 90% što upućuje na genski jako očuvano područje. To pokazuje da je virus Marekove bolesti proširen u Gujaratu
