Development of a molecular-beacon-based multi-allelic real-time RT-PCR assay for the detection of human coronavirus causing severe acute respiratory syndrome (SARS-CoV): A general methodology for detecting rapidly mutating viruses
| Autor: | Hadjinicolaou, Andreas V., Farcas, Gabriella A., Demetriou, Victoria L., Mazzulli, T., Poutanen, Susan M., Willey, B. M., Low, D. E., Butany, Jagdish W., Asa, S. L., Kain, K. C., Kostrikis, Leontios G. |
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| Přispěvatelé: | Kostrikis, Leontios G. [0000-0002-5340-7109] |
| Rok vydání: | 2011 |
| Předmět: |
Severe Acute Respiratory Syndrome
medicine.disease_cause 0302 clinical medicine oligonucleotide probe Porcine Epidemic Diarrhea Virus Coronaviridae Multiplex genetics 030212 general & internal medicine Coronavirus 0303 health sciences education.field_of_study evaluation Reverse Transcriptase Polymerase Chain Reaction article methodology General Medicine SARS Virus 3. Good health virology Reverse transcription polymerase chain reaction Severe acute respiratory syndrome-related coronavirus Molecular Diagnostic Techniques Original Article Severe acute respiratory syndrome SARS coronavirus Population Infectious Bronchitis Virus severe acute respiratory syndrome Biology chemistry Sensitivity and Specificity reverse transcription polymerase chain reaction 03 medical and health sciences Molecular beacon Molecular Beacon Virology molecular diagnosis medicine Humans human education Gene 030304 developmental biology isolation and purification medicine.disease biology.organism_classification sensitivity and specificity Mutation mutation Oligonucleotide Probes |
| Zdroj: | Archives of Virology Arch.Virol. |
| Popis: | Emerging infectious diseases have caused a global effort for development of fast and accurate detection techniques. The rapidly mutating nature of viruses presents a major difficulty, highlighting the need for specific detection of genetically diverse strains. One such infectious agent is SARS-associated coronavirus (SARS-CoV), which emerged in 2003. This study aimed to develop a real-time RT-PCR detection assay specific for SARS-CoV, taking into account its intrinsic polymorphic nature due to genetic drift and recombination and the possibility of continuous and multiple introductions of genetically non-identical strains into the human population, by using mismatch-tolerant molecular beacons designed to specifically detect the SARS-CoV S, E, M and N genes. These were applied in simple, reproducible duplex and multiplex real-time PCR assays on 25 post-mortem samples and constructed RNA controls, and they demonstrated high target detection ability and specificity. This assay can readily be adapted for detection of other emerging and rapidly mutating pathogens. © 2011 Springer-Verlag. 156 671 680 Cited By :5 |
| Databáze: | OpenAIRE |
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