Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/18418
Title: Interlaboratory Reproducibility of Droplet Digital Polymerase Chain Reaction Using a New DNA Reference Material Format.
Authors: Pinheiro, Leonardo B;O'Brien, Helen;Druce, Julian;Do, Hongdo;Kay, Pippa;Daniels, Marissa;You, Jingjing;Burke, Daniel;Griffiths, Kate;Emslie, Kerry R
Affiliation: Research and Development, Australian Red Cross Blood Service , Kelvin Grove, Queensland, Australia
Victorian Infectious Diseases Reference Laboratory , Melbourne, Victoria, Australia
Translational Genomics and Epigenomics Laboratory, Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria, Australia
Agri-Bio Molecular Genetics, Biosciences Research Division, Bundoora, Victoria 3083, Australia
The Prince Charles Hospital University of Queensland , Thoracic Research Centre, Chermside, Queensland, Australia
Save Sight Institute, Sydney Eye Hospital, Sydney Medical School, University of Sydney , Sydney, New South Wales, Australia
National Measurement Institute (NMI) , Lindfield, Sydney, New South Wales, Australia
Issue Date: 7-Nov-2017
EDate: 2017-10-18
Citation: Analytical chemistry 2017; 89(21): 11243-11251
Abstract: Use of droplet digital PCR technology (ddPCR) is expanding rapidly in the diversity of applications and number of users around the world. Access to relatively simple and affordable commercial ddPCR technology has attracted wide interest in use of this technology as a molecular diagnostic tool. For ddPCR to effectively transition to a molecular diagnostic setting requires processes for method validation and verification and demonstration of reproducible instrument performance. In this study, we describe the development and characterization of a DNA reference material (NMI NA008 High GC reference material) comprising a challenging methylated GC-rich DNA template under a novel 96-well microplate format. A scalable process using high precision acoustic dispensing technology was validated to produce the DNA reference material with a certified reference value expressed in amount of DNA molecules per well. An interlaboratory study, conducted using blinded NA008 High GC reference material to assess reproducibility among seven independent laboratories demonstrated less than 4.5% reproducibility relative standard deviation. With the exclusion of one laboratory, laboratories had appropriate technical competency, fully functional instrumentation, and suitable reagents to perform accurate ddPCR based DNA quantification measurements at the time of the study. The study results confirmed that NA008 High GC reference material is fit for the purpose of being used for quality control of ddPCR systems, consumables, instrumentation, and workflow.
URI: http://ahro.austin.org.au/austinjspui/handle/1/18418
DOI: 10.1021/acs.analchem.6b05032
ORCID: 0000-0002-6854-8465
PubMed URL: 28968098
Type: Journal Article
Appears in Collections:Journal articles

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