Caister Academic Press

Internal Amplification Controls in Real-time Polymerase Chain Reaction-Based Methods for Pathogen Detection

Nigel Cook, Gabriel A de Ridder, Martin D'Agostino and Maureen B Taylor
from: Polymerase Chain Reaction: Theory and Technology (Author: Mark A. Behlke, Kornelia Berghof-Jäger, Tom Brown, et al.). Caister Academic Press, U.K. (2019) Pages: 179-186.

Abstract

Assays based on nucleic acid amplification are highly efficient, but they can be affected by the presence of matrix-derived substances which can interfere or prevent the reaction from performing correctly. Careful sample treatment must be applied/used to remove these inhibitory substances. However no sample treatment can be relied on completely, thus an amplification control should be employed to be able to verify that the assay has performed correctly. An internal amplification control (IAC) is a non-target DNA sequence present in the very same reaction as the sample or target nucleic acid extract. If it is successfully amplified to produce a signal, any non-production of a target signal in the reaction is considered to signify that the sample did not contain the target pathogen or organism. If however the reaction produces neither a signal from the target nor the IAC, it signifies that the reaction has failed read more ...
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