Taqman real-time PCR detects Avipoxvirus DNA in blood of Hawai'i 'amakihi (Hemignathus virens).
BACKGROUND
Avipoxvirus sp. is a significant threat to endemic bird populations on several groups of islands worldwide, including Hawai'i, the Galapagos Islands, and the Canary Islands. Accurate identification and genotyping of Avipoxvirus is critical to the study of this disease and how it interacts with other pathogens, but currently available methods rely on invasive sampling of pox-like lesions and may be especially harmful in smaller birds.


RESULTS
Here, we present a nested TaqMan Real-Time PCR for the detection of the Avipoxvirus 4b core protein gene in archived blood samples from Hawaiian birds. The method was successful in amplifying Avipoxvirus DNA from packed blood cells of one of seven Hawaiian honeycreepers with confirmed Avipoxvirus infections and 13 of 28 Hawai'i 'amakihi (Hemignathus virens) with suspected Avipoxvirus infections based on the presence of pox-like lesions. Mixed genotype infections have not previously been documented in Hawai'i but were observed in two individuals in this study.


CONCLUSIONS
We anticipate that this method will be applicable to other closely related strains of Avipoxvirus and will become an important and useful tool in global studies of the epidemiology of Avipoxvirus.
Department of Pharmaceutical Sciences, College of Pharmacy, University of Hawai'i at Hilo, Hilo, Hawai'i, United States of America.
Margaret E M
ME
Farias
Dennis A
DA
LaPointe
Carter T
CT
Atkinson
Christopher
C
Czerwonka
Rajesh
R
Shrestha
Susan I
SI
Jarvi
D000818:Animals; D018150:Avipoxvirus / Q000235:genetics*; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics*; D046109:Passeriformes / Q000097:blood* / Q000821:virology*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D019914:Taq Polymerase / Q000378:metabolism*
D004279:DNA, Viral; D019914:Taq Polymerase