Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6133417 | Journal of Virological Methods | 2015 | 6 Pages |
Abstract
A rapid and accurate method of detection and differentiation of virulent and avirulent Newcastle disease virus (NDV) pathotypes was developed. The NDV detection was carried out for different domestic avian field isolates and pigeon paramyxo virus-1 (25 field isolates and 9 vaccine strains) by using APMV-I “fusion” (F) gene Class II specific external primer A and B (535Â bp), internal primer C and D (238Â bp) based reverses transcriptase PCR (RT-PCR). The internal degenerative reverse primer D is specific for F gene cleavage position of virulent strain of NDV. The nested RT-PCR products of avirulent strains showed two bands (535Â bp and 424Â bp) while virulent strains showed four bands (535Â bp, 424Â bp, 349Â bp and 238Â bp) on agar gel electrophoresis. This is the first report regarding development and use of degenerate primer based nested RT-PCR for accurate detection and differentiation of NDV pathotypes by demonstrating multiple PCR band patterns. Being a rapid, simple, and economical test, the developed method could serve as a valuable alternate diagnostic tool for characterizing NDV isolates and carrying out molecular epidemiological surveillance studies for this important pathogen of poultry.
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Authors
P.A. Desingu, S.D. Singh, K. Dhama, O.R. Vinodh Kumar, R. Singh, R.K. Singh,