Comparison of storage methods for reverse-transcriptase PCR amplification of rotavirus RNA from gorilla ( Gorilla g. gorilla) fecal samples

• Published in: Journal of virological methods 2004-03, Vol.116 (1), p.11-17
• Main Authors: Whittier, Christopher A, Horne, William, Slenning, Barrett, Loomis, Michael, Stoskopf, Michael K
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Desiccation; Ethanol; Fecal; Feces - virology; Formaldehyde; Fundamental and applied biological sciences. Psychology; Gorilla; Gorilla gorilla - virology; Guanidines; Microbiology; Preservation, Biological - methods; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral - analysis; RNA, Viral - isolation & purification; Rotavirus; Rotavirus - genetics; Rotavirus - isolation & purification; rt-PCR; Storage methods; Techniques used in virology; Thiocyanates; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2003.10.003

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.