Correlation between Two RT-PCR Primers in Detection of Avian Influenza Viruses (H9N2)

Document Type : Full Research Paper

Authors

1 Department of Microbiology, College of basic sciences, Shahre Qods branch of Islamic Azad University, Shahre Qods, Iran

2 عضو هیات علمی تمام وقت دانشگاه آزاد اسلامی واحد کرج

3 Department of Clinical Science, College of veterinary medicine, Karaj branch of Islamic Azad University, Karaj, Iran

Abstract

The goal of current study was comparison of two RT-PCR primers in detection of H9N2 influenza viruses. We compare correlation between two methods to determine which method in serial dilution can detect more samples. First, EID50 of an avian influenza virus was calculated by Reed-Muench method. The titer of virus was 106 EID50/ml. Then, we prepare two fold serial dilution of virus. Then, genome was extracted by RNA Pure kit (Cinna-Gene, Iran). Two separate one step RT-PCR methods were used to detection of avian influenza virus (Wu et al. 2008 and Spackman et al. 2002). Target gene of both methods was matrix (M) gene. Product size that produced by both methods were 131 bp and 100 bp respectively. To ensure about specifity of two RT-PCR primers, we used H120 vaccine and B1 vaccine as negative control. Both methods were specific to avian influenza virus and none of them did not produce any amplicons when used other respiratory disease viruses as template. The Wu et al, 2008 primers was able to detect avian influenza virus to 1:4096 and 168 copy number/ml but, Spackman et al, 2000 primers was only able to detect 1:2048 and 336 copy number/ml. So, the Wu et al, 2008 primers was more sensitive method in detection of avian influenza viruses.

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References

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Veterinary Research & Biological Products
Volume 32, Issue 4
December 2019
Pages 38-43

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History

  • Receive Date: 04 September 2018
  • Revise Date: 25 February 2019
  • Accept Date: 26 February 2019

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