Rapid and efficient method to eliminate substances inhibitory to the polymerase chain reaction from animal fecal samples.
To detect pathogenic viruses in animal fecal specimens by polymerase chain reaction (PCR) assays, it is important to remove or inactivate PCR-inhibitory substances. Recently, it was reported that such inhibitory substances in human feces could be efficiently eliminated by a cationic surfactant, Catrimox-14 (Iowa Biotechnology, Iowa) during extraction of viral RNA. In the present report, Catrimox-14 was successfully applied to detect pathogenic viruses in fecal specimens from a variety of animals. By extraction of viral DNA in the presence of this cationic surfactant, the PCR assay could detect canine parvovirus (CPV) in all fecal specimens prepared from 13 kinds of animals, i.e., cat, chicken, cow, dog, gerbil, goat, golden hamster, horse, mouse, pig, rat, rabbit, or sheep. Pretreatment by gel-filtration or boiling failed to remove or inactivate the PCR-inhibitory substances in fecal specimens from mouse, goat, rat, and sheep.
Laboratory of Veterinary Virology, College of Agriculture and Veterinary Medicine, Nihon University, Kanagawa, Japan.
K
K
Uwatoko
M
M
Sunairi
A
A
Yamamoto
M
M
Nakajima
K
K
Yamaura
D000818:Animals; D001483:Base Sequence; D002415:Cats; D002645:Chickens; D006224:Cricetinae; D017931:DNA Primers; D004285:Dogs; D005243:Feces / Q000821:virology*; D005849:Gerbillinae; D006041:Goats; D006736:Horses; D006801:Humans; D008647:Mesocricetus; D051379:Mice; D010865:Pilot Projects; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D000644:Quaternary Ammonium Compounds; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D013501:Surface-Active Agents; D013552:Swine; D050337:Trimethyl Ammonium Compounds; D014780:Viruses / Q000302:isolation & purification*
D017931:DNA Primers; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; D050337:Trimethyl Ammonium Compounds; C009410:tetradecyltrimethylammonium