Comparison of storage methods for reverse-transcriptase PCR amplification of rotavirus RNA from gorilla (Gorilla g. gorilla) fecal samples
| Autor: | Barrett D. Slenning, Michael K. Stoskopf, William A. Horne, Christopher A. Whittier, Michael R. Loomis |
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| Rok vydání: | 2004 |
| Předmět: |
Rotavirus
Preservation Biological Reoviridae medicine.disease_cause Guanidines Virus law.invention Feces law Formaldehyde Virology medicine Animals Desiccation Polymerase chain reaction Gorilla gorilla Ethanol biology Reverse Transcriptase Polymerase Chain Reaction RNA biology.organism_classification Molecular biology Reverse transcription polymerase chain reaction Real-time polymerase chain reaction Nucleic acid RNA Viral Thiocyanates |
| Zdroj: | Journal of Virological Methods. 116:11-17 |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/j.jviromet.2003.10.003 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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