Comprehensive Panel of Real-Time TaqMan™ Polymerase Chain Reaction Assays for Detection and Absolute Quantification of Filoviruses, Arenaviruses, and New World Hantaviruses
Autor: | David A. Kulesh, Leslie Wachter, Valerie A. Buckley-Beason, Randal J. Schoepp, Joseph N. Fair, Jordan B. Jahrling, Jeffrey Garrison, David A. Norwood, Lisa E. Hensley, Adrienne R. Trombley, Augustine Goba |
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Rok vydání: | 2010 |
Předmět: |
Orthohantavirus
Arenavirus biology viruses Filoviridae Articles biology.organism_classification medicine.disease Polymerase Chain Reaction Sensitivity and Specificity Virology Viral hemorrhagic fever Marburg virus Infectious Diseases medicine TaqMan Humans RNA Viral Parasitology Bunyaviridae Viral load Hantavirus |
Zdroj: | The American Journal of Tropical Medicine and Hygiene. 82:954-960 |
ISSN: | 1476-1645 0002-9637 |
Popis: | Viral hemorrhagic fever is caused by a diverse group of single-stranded, negative-sense or positive-sense RNA viruses belonging to the families Filoviridae (Ebola and Marburg), Arenaviridae (Lassa, Junin, Machupo, Sabia, and Guanarito), and Bunyaviridae (hantavirus). Disease characteristics in these families mark each with the potential to be used as a biological threat agent. Because other diseases have similar clinical symptoms, specific laboratory diagnostic tests are necessary to provide the differential diagnosis during outbreaks and for instituting acceptable quarantine procedures. We designed 48 TaqMan™-based polymerase chain reaction (PCR) assays for specific and absolute quantitative detection of multiple hemorrhagic fever viruses. Forty-six assays were determined to be virus-specific, and two were designated as pan assays for Marburg virus. The limit of detection for the assays ranged from 10 to 0.001 plaque-forming units (PFU)/PCR. Although these real-time hemorrhagic fever virus assays are qualitative (presence of target), they are also quantitative (measure a single DNA/RNA target sequence in an unknown sample and express the final results as an absolute value (e.g., viral load, PFUs, or copies/mL) on the basis of concentration of standard samples and can be used in viral load, vaccine, and antiviral drug studies. |
Databáze: | OpenAIRE |
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