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Review
. 2017 Feb 2:8:108.
doi: 10.3389/fmicb.2017.00108. eCollection 2017.

A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything

Petr Kralik  1 , Matteo Ricchi  2
Affiliations
Review

A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything

Petr Kralik et al. Front Microbiol. .

Abstract

Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the areas of clinical and veterinary diagnostics and food safety. Although the concept of PCR is relatively simple, there are specific issues in qPCR that developers and users of this technology must bear in mind. These include the use of correct terminology and definitions, understanding of the principle of PCR, difficulties with interpretation and presentation of data, the limitations of qPCR in different areas of microbial diagnostics and parameters important for the description of qPCR performance. It is not our intention in this review to describe every single aspect of qPCR design, optimization, and validation; however, it is our hope that this basic guide will help to orient beginners and users of qPCR in the use of this powerful technique.

Keywords: accuracy; limit of detection; limit of quantification; precision; quantitative PCR; trueness.

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Figures

Figure 1
Figure 1
Model calibration curve with the regression equation (characterized by the slope and intercept) and regression coefficient.
Figure 2
Figure 2
Graphical representation of the determination of limit of detection (LOD) in qPCR. The data in the table show the number of positive samples/all analyzed samples (signal ratio).
See this image and copyright information in PMC

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