Assessment of the sensitivity of Real-time PCR for diagnosis of Newcastle disease virus

Document Type : Full Research Paper

Authors

1 M.Sc student of animal genetics and breeding, Faculty of Agriculture, Razi University, Kermanshah, Iran.

2 Member of scientific board, Department of Biotechnology, Animal Science Research Institute of Iran (ASRI), Karaj, Iran.

3 Member of scientific board, Faculty of Agriculture, Razi University, Kermanshah, Iran.

4 Member of scientific board, Nanosystems Research team, Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran.

Abstract

Newcastle disease is one of the most serious viral diseases in the poultry worldwide. Since the traditional strategies have been not control it well, the aim of this study was to introduce new methods for early and rapid diagnosis of Newcastle. In this study, to detect the virus, a Real-time PCR method was optimized,Viral RNA was extracted from strain B1 using the kit RNease mini (Qiagen, USA), according to the manufacturer's instructions. This sample had 68.23×109copy numbers of viral RNA per each microliter. Then, a serial dilution as 100-fold was prepare from the initial sample. Then, these dilutions were simultaneously applied in reverse transcription and Realtime PCR using commercial kits (Genekam Biotechnology, Germany) according to the manufacture’s instruction. The sensitivity of Real-time PCR reaction was determined based on serial dilutions of 1×10-34 copy number per micro liter. Since, speed and accuracy in diagnosis of contagious of Newcastle disease virus plays an important role to control the disease, so adopting this method is recommended as a diagnostic test with high sensitivity.

Keywords

References

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History

  • Receive Date: 22 May 2015
  • Revise Date: 31 January 2016
  • Accept Date: 23 September 2015

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