Quality control for quantitative PCR based on amplification compatibility test

0351162 - BTÚ 2011 RIV US eng J - Journal Article
Tichopád, Aleš - Bar, T. - Pecen, Ladislav - Kitchen, R.R. - Kubista, Mikael - Pfaffl, M.W.
Quality control for quantitative PCR based on amplification compatibility test.
Methods. Roč. 50, č. 4 (2010), s. 308-312. ISSN 1046-2023. E-ISSN 1095-9130
R&D Projects: GA AV ČR IAA500520809; GA AV ČR IAA500970904
Institutional research plan: CEZ:AV0Z50520701
Keywords : Quantitative PCR * Quality control * Amplification efficiency
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 4.527, year: 2010

Quantitative qPCR is method for the accurate quantification of nucleic acids. Yet it may generate erroneous results if the amplification process is obscured by inhibition or generation of aberrant side-product. Herein we present a statistical approach based on multivariate analysis of the amplification response data generated in qPCR. The amplification trajectory in its most dynamic phase is fitted with a suitable model. Two parameters of this model, related to amplification efficiency, are then used for calculation of the Z-score statistics. Each studied sample is compared to a predefined reference set of reactions. A probabilistic decision for each individual Z-score is then used to identify the majority of inhibited reactions in our experiments. We demonstrate improved identification performance using the multivariate approach Finally we stress that the performance of the amplification kompatibility test as a quality control procedure depends on the quality of the reference set
Permanent Link: http://hdl.handle.net/11104/0190971