In-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory viruses
<b>Background</b>. Multiplex real time PCR is increasingly used to diagnose respiratory viruses and has shown to be superior to traditional methods, such as culture and antigen detection. <b>Objective</b>. Standardization and validation of a multiplex real-time PCR assay for...
| Autores: | , , , , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2016 |
| País: | Argentina |
| Institución: | Universidad Nacional de La Plata |
| Repositorio: | SEDICI (UNLP) |
| Idioma: | inglés |
| OAI Identifier: | oai:sedici.unlp.edu.ar:10915/122375 |
| Acceso en línea: | http://sedici.unlp.edu.ar/handle/10915/122375 |
| Access Level: | acceso abierto |
| Palabra clave: | Veterinaria Multiplex Real Time PCR Respiratory virus Standardization PRC múltiple en tiempo real virus respiratorio estandarización |
| Sumario: | <b>Background</b>. Multiplex real time PCR is increasingly used to diagnose respiratory viruses and has shown to be superior to traditional methods, such as culture and antigen detection. <b>Objective</b>. Standardization and validation of a multiplex real-time PCR assay for the detection of 13 respiratory viruses. <b>Methods</b>. The assay was validated using RNA control targets and comparing results to single-target PCR’s. <b>Results</b>. Using RNA controls the multiplex format was found to be as sensitive and specific as the single-target PCRs, and no competition was observed between targets. The efficiencies for most of the reactions were approximately 90%, but a lower efficiency was found for Parainfluenza 2 with a rate of amplification in each cycle of 86.63%. On the other hand, a higher efficiency was observed in respiratory syncytial virus A and respiratory syncytial virus B ((93.07% each). <b>Conclusion</b>: This multiplex RT-PCR format shows an adequate efficiency, demonstrating an excellent sensitivity, specificity and repeatability for all the studied respiratory viruses. |
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