Evidence map›Paper›PMID 32640980›Full record

ArticleBMC bioinformatics2020

Critique of the pairwise method for estimating qPCR amplification efficiency: beware of correlated data!

Joel Tellinghuisen

Abstract read
In one paragraph

Article in BMC bioinformatics, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed
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1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

1 author.

Joel TellinghuisenDepartment of Chemistry, Vanderbilt University, Nashville, TN, 37235, USA. joel.tellinghuisen@vanderbilt.edu.ORCID http://orcid.org/0000-0002-4487-5197

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundA recently proposed method for estimating qPCR amplification efficiency E analyzes fluorescence intensity ratios from pairs of points deemed to lie in the exponential growth region on the amplification curves for all reactions in a dilution series. This method suffers from a serious problem: The resulting ratios are highly correlated, as they involve multiple use of the raw data, for example, yielding ~ 250 E estimates from ~ 25 intensity readings. The resulting statistics for such estimates are falsely optimistic in their assessment of the estimation precision.

resultsMonte Carlo simulations confirm that the correlated pairs method yields precision estimates that are better than actual by a factor of two or more. This result is further supported by estimating E by both pairwise and C

conclusionContrary to assertions in the proposing work, the pairwise method does not yield E estimates a factor of 2 more precise than estimates from C

Indexed as

Real-Time Polymerase Chain ReactionCorrelation of DataFluorescenceMonte Carlo MethodAmplification efficiencyCalibrationCorrelated dataData analysisqPCRStatistical errorsWeighted least squares

Identifiers

PMID32640980
PMCPMC7346608

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.