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

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

dc.creatorVargas, Hernán
dc.creatorDiaz, Ángela
dc.creatorCelis, Yamile
dc.creatorDíaz, Liliana
dc.creatorGómez, Sandra
dc.creatorSánchez, Jenny
dc.creatorGolijow, Carlos
dc.creatorArce, Patricia
dc.date2016-12-15
dc.date.accessioned2022-02-22T19:57:51Z
dc.date.available2022-02-22T19:57:51Z
dc.identifierhttps://hemeroteca.unad.edu.co/index.php/nova/article/view/1746
dc.identifier10.22490/24629448.1746
dc.identifier.urihttps://repository.unad.edu.co/handle/10596/46876
dc.descriptionBackground. 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.en-US
dc.descriptionBackground. 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.es-ES
dc.formatapplication/pdf
dc.languageeng
dc.publisherUniversidad Colegio Mayor de Cundinamarcaes-ES
dc.relationhttps://hemeroteca.unad.edu.co/index.php/nova/article/view/1746/1988
dc.rightsDerechos de autor 2017 NOVA Publicación en Ciencias Biomédicases-ES
dc.sourceNOVA Biomedical Sciences Journal; Vol. 14 No. 26 (2016); 9-15en-US
dc.sourceNova; Vol. 14 Núm. 26 (2016); 9-15es-ES
dc.sourceNOVA Ciências Biomédicas Publicação; v. 14 n. 26 (2016); 9-15pt-BR
dc.source2462-9448
dc.source1794-2470
dc.subjectMultiplex Real Time PCRen-US
dc.subjectRespiratory virusen-US
dc.subjectStandardizationen-US
dc.subjectMultiplex Real Time PCRes-ES
dc.subjectRespiratory viruses-ES
dc.subjectStandardization.es-ES
dc.titleIn-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory virusesen-US
dc.titleIn-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory viruseses-ES
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:eu-repo/semantics/publishedVersion
dc.typeinfo:eu-repo/article/publisheden-US
dc.typeinfo:eu-repo/article/publishedes-ES
dc.typeinfo:eu-repo/article/publishedpt-BR


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