Real‐time NASBA detection of SARS‐associated coronavirus and comparison with real‐time reverse transcription‐PCR
| Autor: | Peter Sillekens, Maria-Cristina Keightley, Charles R. Rinaldo, Kirsten St. George, Wim Schippers |
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| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
RT‐PCR
viruses coronavirus medicine.disease_cause Severe Acute Respiratory Syndrome Article NASBA Virology medicine Coronaviridae Humans skin and connective tissue diseases Polymerase Coronavirus SARS biology Reverse Transcriptase Polymerase Chain Reaction fungi RNA biology.organism_classification Reverse transcription polymerase chain reaction Infectious Diseases Real-time polymerase chain reaction Severe acute respiratory syndrome-related coronavirus biology.protein RNA Viral Primer (molecular biology) Nucleic Acid Amplification Techniques Research Article |
| Zdroj: | Journal of Medical Virology |
| ISSN: | 1096-9071 0146-6615 |
| Popis: | Severe acute respiratory syndrome (SARS) exhibits a high mortality rate and the potential for rapid epidemic spread. Additionally, it has a poorly defined clinical presentation, and no known treatment or prevention methods. Collectively, these factors underscore the need for early diagnosis. Molecular tests have been developed to detect SARS coronavirus (SARS‐CoV) RNA using real time reverse transcription polymerase chain reaction (RT‐PCR) with varying levels of sensitivity. However, RNA amplification methods have been demonstrated to be more sensitive for the detection of some RNA viruses. We therefore developed a real‐time nucleic acid sequence‐based amplification (NASBA) test for SARS‐CoV. A number of primer/beacon sets were designed to target different regions of the SARS‐CoV genome, and were tested for sensitivity and specificity. The performance of the assays was compared with RT‐PCR assays. A multi‐target real‐time NASBA application was developed for detection of SARS‐CoV polymerase (Pol) and nucleocapsid (N) genes. The N targets were found to be consistently more sensitive than the Pol targets, and the real‐time NASBA assay demonstrates equivalent sensitivity when compared to testing by real‐time RT‐PCR. A multi‐target real‐time NASBA assay has been successfully developed for the sensitive detection of SARS‐CoV. J. Med. Virol. 77:602–608, 2005. © 2005 Wiley‐Liss, inc. |
| Databáze: | OpenAIRE |
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