What is it about?
MaxRatio is a mathematical algorithm recently introduced that, unlike the 'cycle-threshold' commonly used for the analysis of the qPCR data, does not rely upon a threshold defined by the operator. We apply MaxRatio to the qPCR data generated by the platform TaqMan Array Card by ThermoFisher that used 48 different primer sets. We observed that there was an increase in the accuracy of the results in 20% of the primer sets in comparison to the standard 'cycle-threshold' method.
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Why is it important?
Just by changing the analytical algorithm, 1/5 of the primer sets in use showed an increase in accuracy. Although marginal, this increase might be useful in the clinical setting where thousands of PCR reactions are validated monthly. In particular, the negative predictive value was increased using MaxRatio, so there was a higher probability of issuing truly negative results than by using the 'cycle-threshold'.
Perspectives
MaxRatio can be easily implemented in computational pipelines that can automate the analysis of the qPCR data relieving the operator from visually validating large amounts of reactions.
Luigi Marongiu
Read the Original
This page is a summary of: Assumption-free improvement of the maxRatio algorithm increases the accuracy of qPCR assays targeting viruses, Journal of Clinical Virology, September 2016, Elsevier,
DOI: 10.1016/j.jcv.2016.08.011.
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