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The applicability of digital PCR for the assessment of detection limits in GMO analysis

Burns, M.J., Burrell, A.M., Foy, C.A.

We investigate the applicability of using digital PCR to estimate absolute limits of detection and quantitation associated with the validation of traditional real-time PCR methods for analysis of genetically modified (GM) ingredients. We also demonstrate the use of dynamic arrays as a precursor in selecting suitable sample dilution levels for accurate copy number assessment using the aforementioned digital PCR. Additionally, we further explore the relevance of digital PCR in accurately quantifying plasmid copy numbers associated with a commercially available Certified Reference Material. The use of digital PCR has the advantage of facilitating absolute single molecule detection, therefore negating the necessity for standards on a calibration curve, and reducing associated matrix effects. The approaches described in this paper enable pre-existing validated protocols to be re-examined, and estimates based on an alternative approach using digital PCR to be used in order to objectively characterise sensitivity limits.

Citation

Burns, M.J., Burrell, A.M., Foy, C.A. "The applicability of digital PCR for the assessment of detection limits in GMO analysis" European Food Research and Technology (2010): 353–62