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Publication Development of Molecular Diagnostic Methods for Detecting West Nile ...  

Update Date: [2013-11-14]

NO.: 42
 
Reportno. AHRI report No.42
Topic

Development of Molecular Diagnostic Methods for Detecting West Nile Virus in Wild Birds in Taiwan in 2006

Department

1Animal Health Research Institute, Council of Agriculture, Executive Yuan
2 Centers of Disease Control and Prevention, United States

Author

Liu YP1*1, Chang GJ2, Cheng MC1, Lee MS1, Chen LH1, Lee SH1

Summary
West Nile virus (WNV) infection is an important zoonosis. To prevent the possible introduction of this emerging disease into Taiwan , we have established a comprehensive wild-bird surveillance system for monitoring the presence of West Nile viral RNAs in migratory birds. Three detection methods for West Nile viral RNA, including nested PCR, SYBR Green I real-time PCR and Multiplex TaqMan real-time PCR, were used for this study. The nested PCR assay could detect 5 RNA copies, and it was 1000-fold more sensitive than that of traditional PCR. The sensitivity of SYBR Green I real-time PCR assay was equal to nested PCR. However, if the standard curve generated from known copy-number control specimens was included in the assay, the SYBR Green assay could be used to quantify the copy number of testing specimens. The multiplex TaqMan real-time PCR could differentiate four important flaviviruses, including WNV, JE, SLE and YF viruses. We have incorporated the surveillance project of WNV into the avian influenza surveillance project established previously. A total of 4,626 specimens from wild birds were processed in 2006, and none of them was WNV positive 

Keyword

West Nile fever, Wild birds, Real-time polymerase chain reaction