Rapid Detection/pathotyping of Newcastle disease virus isolates in clinical samples using real time polymerase chain reaction assay
Abdul wajid kakar, Abdul Wajida, Muhammad Wasima, Tahir Yaqubb, Shafqat Fatima Rehmanib, Tasra Bibic, Nadia Mukhtard, Javed Muhammad, Umar Bachae, Suliman Qadir Afridi, Muhammad Nauman Zahid, Zia u ddin, Muhammad Zubair Shabbirb, Kamran Abbasa, Munir Ahmed
- aInstitute of Biochemistry and Biotechnology , University of Veterinary and Animal Sciences Lahore 54000 , Pakistan
- bQuality Operations Laboratory , University of Veterinary and Animal Sciences Lahore 54000 , Pakistan
- cDepartment of Pathology , University of Veterinary and Animal Sciences Lahore 54000 , Pakistan
- dDepartment of Microbiology , University of Veterinary and Animal Sciences Lahore 54000 , Pakistan Javed Muhammad University Diagnostic Lab (UDL)
- eDepartment of Food Science and human nutrition , University of Veterinary and Animal Sciences Lahore 54000 , Pakistan
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Abstract
In the present protocol we describe the real time reverse transcription polymerase chain reaction \(rRT-PCR) assay for the rapid detection/pathotyping of Newcastle disease virus \(NDV) isoaltes in clinical samples.Fusion gene and matrix gene assays have been used to rapidly detect the NDV isolates.The F gene assay is designed used degenerated oligonucleotide primers and TaqMan probes with non uorescent minor groove binder \(MGB) quencher to differentiate lentogenic \(low virulent) and velogenic \(high virulent) NDVs based on cleavage site of the F0 precursor of fusion protein gene which is a key determinant of NDV pathogenicity.