Evaluation of real-time PCR results at the limit of detection

被引：0

作者：

Waiblinger, Hans-Ulrich ^{[1
]}

Graf, Norbert ^{[2
]}

Broll, Hermann ^{[3
]}

Grohmann, Lutz ^{[4
]}

Pietsch, Klaus ^{[1
]}

机构：

[1] Chem & Veterinaruntersuchungsamt Freiburg State I, D-79114 Freiburg, Germany

[2] Inst Lebensmittelchem Trier, Landesuntersuchungsamt Rheinland Pfalz, D-54295 Trier, Germany

[3] Bundesinst Risikobewertung, D-14195 Berlin, Germany

[4] Bundesamt Verbraucherschutz & Lebensmittelsicherh, D-10117 Berlin, Germany

来源：

JOURNAL FUR VERBRAUCHERSCHUTZ UND LEBENSMITTELSICHERHEIT-JOURNAL OF CONSUMER PROTECTION AND FOOD SAFETY | 2011年 / 6卷 / 04期

关键词：

Real-time PCR; Limit of detection; GMO; Allergens; Zero tolerance;

D O I：

10.1007/s00003-011-0669-4

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

Real-time polymerase chain reaction (real-time PCR) facilitates to detect DNA fragments at very low copy numbers. Positive results, e.g. for unauthorized genetically modified organism (GMO) contamination or for allergens, may raise safety concerns and have far-reaching consequences. However, in case of very low concentrations of DNA samples, results for the same product lot or even for identical samples from different laboratories may differ. Therefore, an approach for a standardized interpretation and reporting of results obtained by real-time PCR at the limit of detection (LOD) is proposed. A quality control DNA sample containing the target at the LOD (95 %) is analysed in parallel with the real DNA sample and the respective C (T) values are compared. In addition, practical approaches for in house and precision-based estimation of the LOD are presented. The proposed approach may also contribute to the current discussion on implementing a technical solution to handle DNA traces in specimen, e.g. for the detection of unauthorized GMO.

引用

页码：411 / 417

页数：7

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