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Montag, 8. Mai 2017
Methods to determine limit of detection and limit of quantification in qPCR
In Press, Corrected Proof, Available online 29 April 2017
Amin Forootan, Robert Sjöback, Jens Björkman, Björn Sjögreen, Lucas Linz, Mikael Kubista
Abstract: Quantitative Real-Time Polymerase Chain Reaction, better known as qPCR, is the most sensitive and specific technique we have for the detection of nucleic acids. Even though it has been around for more than 30 years and is preferred in research applications, it has yet to win broad acceptance in routine practice. This requires a means to unambiguously assess the performance of specific qPCR analyses. Here we present methods to determine the limit of detection (LoD) and the limit of quantification (LoQ) as applicable to qPCR. These are based on standard statistical methods as recommended by regulatory bodies adapted to qPCR and complemented with a novel approach to estimate the precision of LoD. Keywords: Limit of detection; Limit of quantification; LoD; LoQ; GenEx software; Real-time PCR; qPCR; Data analysis; Replicates; MIQE; Quality control; Standardization