Document Detail

Validation of PCR methods for quantitation of genetically modified plants in food.
MedLine Citation:
PMID:  11767156     Owner:  NLM     Status:  MEDLINE    
For enforcement of the recently introduced labeling threshold for genetically modified organisms (GMOs) in food ingredients, quantitative detection methods such as quantitative competitive (QC-PCR) and real-time PCR are applied by official food control laboratories. The experiences of 3 European food control laboratories in validating such methods were compared to describe realistic performance characteristics of quantitative PCR detection methods. The limit of quantitation (LOQ) of GMO-specific, real-time PCR was experimentally determined to reach 30-50 target molecules, which is close to theoretical prediction. Starting PCR with 200 ng genomic plant DNA, the LOQ depends primarily on the genome size of the target plant and ranges from 0.02% for rice to 0.7% for wheat. The precision of quantitative PCR detection methods, expressed as relative standard deviation (RSD), varied from 10 to 30%. Using Bt176 corn containing test samples and applying Bt176 specific QC-PCR, mean values deviated from true values by -7to 18%, with an average of 2+/-10%. Ruggedness of real-time PCR detection methods was assessed in an interlaboratory study analyzing commercial, homogeneous food samples. Roundup Ready soybean DNA contents were determined in the range of 0.3 to 36%, relative to soybean DNA, with RSDs of about 25%. Taking the precision of quantitative PCR detection methods into account, suitable sample plans and sample sizes for GMO analysis are suggested. Because quantitative GMO detection methods measure GMO contents of samples in relation to reference material (calibrants), high priority must be given to international agreements and standardization on certified reference materials.
P Hübner; H U Waiblinger; K Pietsch; P Brodmann
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Publication Detail:
Type:  Journal Article; Validation Studies    
Journal Detail:
Title:  Journal of AOAC International     Volume:  84     ISSN:  1060-3271     ISO Abbreviation:  J AOAC Int     Publication Date:    2001 Nov-Dec
Date Detail:
Created Date:  2001-12-19     Completed Date:  2002-06-10     Revised Date:  2008-03-17    
Medline Journal Info:
Nlm Unique ID:  9215446     Medline TA:  J AOAC Int     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1855-64     Citation Subset:  IM    
Kantonales Labor Zürich, Switzerland.
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MeSH Terms
European Union
Food Analysis / methods*,  standards,  statistics & numerical data
Food, Genetically Modified*
Polymerase Chain Reaction / methods*,  standards,  statistics & numerical data
Reference Standards
Sensitivity and Specificity
Zea mays / chemistry,  genetics

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