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Evaluation of different machines used to quantify genetic modification by real-time PCR

Allnutt, Theodore R., Ayadi, Mira, Berben, Gilbert, Brodmann, Peter and Lee, David 2010, Evaluation of different machines used to quantify genetic modification by real-time PCR, Journal of AOAC international, vol. 93, no. 4, pp. 1243-1248.

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Title Evaluation of different machines used to quantify genetic modification by real-time PCR
Author(s) Allnutt, Theodore R.
Ayadi, Mira
Berben, Gilbert
Brodmann, Peter
Lee, David
Journal name Journal of AOAC international
Volume number 93
Issue number 4
Start page 1243
End page 1248
Total pages 6
Publisher AOAC International
Place of publication Rockville, Md.
Publication date 2010-07
ISSN 1060-3271
Keyword(s) Polymerase Chain Reaction
Summary Quantification of genetic modification (GM) is often undertaken to test for compliance with the European Union GM labeling threshold in food. Different control laboratories will often use common validated methods, but with different models of real-time PCR machines. We performed two separate ring trials to evaluate the relative precision and accuracy of different types of real-time PCR machines used to quantify the concentration of GM maize. Both trials used dual-labeled fluorogenic probes for quantification. The first ring trial used separate GM and reference assays (a single fluorescence channel), and the second used a combined duplex assay (two simultaneous fluorescence channels). Five manufacturers and seven models--including a 96-well microtiter-plate, rotary, and portable machines--were examined. In one trial, the machine used had a significant effect on precision, but in the other it did not. Overall, the degree of variation due to the machine model was lower than other factors. No significant repeatable difference in accuracy was observed between machine models. It was not possible to use sufficient replication of machine type in each laboratory to examine all sources of variation in this study, but the results strongly indicate that factors other than machine type or manufacturer (e.g., method or laboratory) contribute more to variation in a GM quantification result.
Language eng
Field of Research 060499 Genetics not elsewhere classified
0301 Analytical Chemistry
0908 Food Sciences
Socio Economic Objective 920406 Food Safety
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2010, AOAC International
Persistent URL http://hdl.handle.net/10536/DRO/DU:30086585

Document type: Journal Article
Collection: School of Medicine
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