Development of Field-Based Real-Time Reverse Transcription–Polymerase Chain Reaction Assays for Detection of Chikungunya and O’nyong-nyong Viruses in Mosquitoes
| Autor: | Jordan B. Jahrling, David A. Kulesh, John S. Lee, Darci R. Smith, Jennifer L. Groebner, Monica L. O'Guinn, Michael J. Turell |
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| Rok vydání: | 2009 |
| Předmět: |
Reverse Transcriptase Polymerase Chain Reaction
RNA Biology medicine.disease_cause Sensitivity and Specificity Virology Reverse transcriptase Virus law.invention Reverse transcription polymerase chain reaction Culicidae Infectious Diseases law medicine Animals RNA Viral Parasitology Chikungunya Viral disease Primer (molecular biology) Chikungunya virus Polymerase chain reaction |
| Zdroj: | The American Journal of Tropical Medicine and Hygiene. 81:679-684 |
| ISSN: | 1476-1645 0002-9637 |
| Popis: | Chikungunya (CHIK) and O'nyong-nyong (ONN) are important emerging arthropod-borne diseases. Molecular diagnosis of these two viruses in mosquitoes has not been evaluated, and the effects of extraneous mosquito tissue on assay performance have not been tested. Additionally, no real-time reverse transcription-polymerase chain reaction (RT-PCR) assay exists for detecting ONN virus (ONNV) RNA. We describe the development of sensitive and specific real-time RT-PCR assays for detecting CHIK and ONN viral RNA in mosquitoes, which have application for field use. In addition, we compared three methods for primer/probe design for assay development by evaluating their sensitivity and specificity. This comparison resulted in development of virus-specific assays that could detect less than one plaque-forming unit equivalent of each of the viruses in mosquitoes. The use of these assays will aid in arthropod-borne disease surveillance and in the control of the associated diseases. |
| Databáze: | OpenAIRE |
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