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Absolute quantification by droplet digital PCR versus analog real-time PCR

Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCR revealed greater precision (coefficients of variation decreased by 37–86%) and improved day-...

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Detalles Bibliográficos
Autores principales: Hindson, Christopher M, Chevillet, John R, Briggs, Hilary A, Gallichotte, Emily N, Ruf, Ingrid K, Hindson, Benjamin J, Vessella, Robert L, Tewari, Muneesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4118677/
https://www.ncbi.nlm.nih.gov/pubmed/23995387
http://dx.doi.org/10.1038/nmeth.2633
Descripción
Sumario:Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCR revealed greater precision (coefficients of variation decreased by 37–86%) and improved day-to-day reproducibility (by a factor of seven) of ddPCR but with comparable sensitivity. When we applied ddPCR to serum microRNA biomarker analysis, this translated to superior diagnostic performance for identifying individuals with cancer.