Comparison of storage methods for reverse-transcriptase PCR amplification of rotavirus RNA from gorilla (Gorilla g. gorilla) fecal samples.
Detection of enteric viral nucleic acids in preserved gorilla fecal specimens was investigated using reverse transcription polymerase chain reaction (rt-PCR). A commercially available viral RNA extraction kit was used to isolate nucleic acids from captive gorilla fecal samples seeded with rotavirus and stored in ethanol, formalin, a commercial RNA preservation solution, guanidine thiocyanate buffer (GT), and samples dried in tubes containing silica gel. Nucleic acids were extracted at 1, 7, 70 and 180 days and used for rt-PCR amplification of specific rotavirus RNA sequences. Successful rt-PCR amplification of the target product varied according to storage conditions, and storage time. Only samples stored in GT gave 100% positive results at 180 days. It is recommended that fecal samples be collected in GT for viral RNA analysis.
Whittier
Christopher A
CA
Environmental Medicine Consortium, College of Veterinary Medicine, North Carolina State University, 4700 Hillsborough Street, Raleigh, NC 27606, USA. chris-whitter@hotmail.com
Horne
William
W
Slenning
Barrett
B
Loomis
Michael
M
Stoskopf
Michael K
MK
D000818:Animals; D003890:Desiccation; D000431:Ethanol; D005243:Feces / Q000821:virology*; D005557:Formaldehyde; D006071:Gorilla gorilla / Q000821:virology*; D006146:Guanidines; D011309:Preservation, Biological / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000235:genetics* / Q000302:isolation & purification; D013861:Thiocyanates
D006146:Guanidines; D012367:RNA, Viral; D013861:Thiocyanates; D005557:Formaldehyde; D000431:Ethanol; C054436:guanidine thiocyanate