Effect of preservative on recoverable RT-PCR amplicon length from influenza A virus in bird feces.
Surveillance for avian influenza A viruses in wild bird populations is often limited by requirements for a cold chain from time of specimen collection, by availability of ultra-low temperature specimen storage within a few hours or days, and by laborious classical virologic procedures. Successful storage of specimens in preservatives at ambient temperature and subsequent detection of RNA by reverse transcriptase-polymerase chain reaction (RT-PCR) would assist in helping influenza surveillance efforts become more widespread in remote areas, as well as more timely and inexpensive. Here, we describe bird feces spiked with influenza A virus preserved with guanidine and commercial buffers or alcohols at ambient temperature and analyzed by RT-PCR protocols. Virus-specific RT-PCR products of, at most, 206 bp were recovered for samples preserved with alcohols and up to 521 bp for samples preserved with guanidine or commercial buffers. These results suggest that this approach is feasible in the field and that preserved specimens might be better assayed molecularly when preserved in guanidine or commercial buffers.
Evers
David L
DL
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Slemons
Richard D
RD
Taubenberger
Jeffery K
JK
D000818:Animals; D002021:Buffers; D004372:Ducks; D005243:Feces / Q000821:virology*; D005777:Geese; D019791:Guanidine / Q000737:chemistry*; D009980:Influenza A virus / Q000302:isolation & purification*; D012367:RNA, Viral; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D013048:Specimen Handling / Q000662:veterinary*
D002021:Buffers; D012367:RNA, Viral; D019791:Guanidine