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Estimating the Accuracy of Polymerase Chain Reaction–Based Tests Using Endpoint Dilution
Author(s) -
Hughes James P.,
Totten Patricia
Publication year - 2003
Publication title -
biometrics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.298
H-Index - 130
eISSN - 1541-0420
pISSN - 0006-341X
DOI - 10.1111/1541-0420.00060
Subject(s) - replicate , nonparametric statistics , sensitivity (control systems) , polymerase chain reaction , parametric statistics , statistics , computational biology , biological system , mathematics , biology , computer science , genetics , gene , electronic engineering , engineering
Summary . Polymerase chain reaction (PCR)–based tests for various microorganisms or target DNA sequences are generally acknowledged to be highly “sensitive,” yet the concept of sensitivity is ill‐defined in the literature on these tests. We propose that sensitivity should be expressed as a function of the number of target DNA molecules in the sample (or specificity, when the target number is 0). However, estimating this “sensitivity curve” is problematic, since it is difficult to construct samples with a fixed number of targets. Nonetheless, using serially diluted replicate aliquots of a known concentration of the target DNA sequence, we show that it is possible to disentangle random variations in the number of target DNA molecules from the underlying test sensitivity. We develop parametric, nonparametric, and semiparametric (spline‐based) models for the sensitivity curve. The methods are compared on a new test for M. genitalium .