In-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory viruses

Abstract

<p>Background. 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. Objective. Standardization and validation of a multiplex real-time PCR assay for the detection of 13 respiratory viruses. Methods. The assay was validated using RNA control targets and comparing results to single-target PCR’s. Results. 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). Conclusion: This multiplex RT-PCR format shows an adequate efficiency, demonstrating an excellent sensitivity, specificity and repeatability for all the studied respiratory viruses.</p>

Análisis de las temáticas y enfoques de los artículos publicados en la Revista NOVA en el periodo comprendido entre 2014 y 2019

“…Por otra parte, técnicas de biología molecular como PCR múltiple en tiempo real y RT-PCR cobran importancia en el análisis de genes relacionados con resistencia bacteriana y son utilizados cada vez más para el diagnóstico de virus respiratorios, mostrando ser métodos superiores en comparación a métodos tradicionales, como cultivo y detección de antígenos (5).…”