title: Absolute quantification by droplet digital PCR versus analog real-time PCR.
abstract: 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 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.
authors.affiliation: 1] Digital Biology Center, Bio-Rad Laboratories, Inc., Pleasanton, California, USA. [2] [3].
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Hindson
authors.forename: John R
authors.initials: JR
authors.lastname: Chevillet
authors.forename: Hilary A
authors.initials: HA
authors.lastname: Briggs
authors.forename: Emily N
authors.initials: EN
authors.lastname: Gallichotte
authors.forename: Ingrid K
authors.initials: IK
authors.lastname: Ruf
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Hindson
authors.forename: Robert L
authors.initials: RL
authors.lastname: Vessella
authors.forename: Muneesh
authors.initials: M
authors.lastname: Tewari
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis* / Q000097:blood; D006801:Humans; D057230:Limit of Detection; D008297:Male; D035683:MicroRNAs / Q000032:analysis* / Q000097:blood; D011471:Prostatic Neoplasms / Q000097:blood / Q000175:diagnosis; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; C533895:MIRN141 microRNA, human; D035683:MicroRNAs