20.500.12556/DiRROS-20035 Comparison and transfer testing of multiplex ligation detection methods for GM plants Background With the increasing number of GMOs on the global market the maintenance of European GMO regulations is becoming more complex. For the analysis of a single food or feed sample it is necessary to assess the sample for the presence of many GMO-targets simultaneously at a sensitive level. Several methods have been published regarding DNA-based multidetection. Multiplex ligation detection methods have been described that use the same basic approach: i) hybridisation and ligation of specific probes, ii) amplification of the ligated probes and iii) detection and identification of the amplified products. Despite they all have this same basis, the published ligation methods differ radically. The present study investigated with real-time PCR whether these different ligation methods have any influence on the performance of the probes. Sensitivity and the specificity of the padlock probes (PLPs) with the ligation protocol with the best performance were also tested and the selected method was initially validated in a laboratory exchange study. Results Of the ligation protocols tested in this study, the best results were obtained with the PPLMD I and PPLMD II protocols and no consistent differences between these two protocols were observed. Both protocols are based on padlock probe ligation combined with microarray detection. Twenty PLPs were tested for specificity and the best probes were subjected to further evaluation. Up to 13 targets were detected specifically and simultaneously. During the interlaboratory exchange study similar results were achieved by the two participating institutes (NIB, Slovenia, and RIKILT, the Netherlands). Conclusions From the comparison of ligation protocols it can be concluded that two protocols perform equally well on the basis of the selected set of PLPs. Using the most ideal parameters the multiplicity of one of the methods was tested and 13 targets were successfully and specifically detected. In the interlaboratory exchange study it was shown that the selected method meets the 0.1% sensitivity criterion. The present study thus shows that specific and sensitive multidetection of GMO targets is now feasible. geneticallz modified organisms GMO true false true Angleški jezik Ni določen Neznano 2024-08-05 07:50:03 2024-08-05 07:50:03 2024-08-07 03:37:38 0000-00-00 00:00:00 2012 0 0 Soavtorji: Jeroen P. van Dijk, Theo W. Prins, Marleen M. Voorhuijzen, A. M. Angeline Van Hoef, Henriek G. Beenen, Dany Morisset, Kristina Gruden and Esther J. Kok; Popis vira z dne 7.2.2012; str. 1-12 no. 4 Vol. 12 2012 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2012-01-12 604 1472-6750 10.1186/1472-6750-12-4 2503503 http://dx.doi.org/10.1186/1472-6750-12-4 1 e911d25e-52dd-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27897 RAZ_Ujhelyi_Gabriella_2012.pdf RAZ_Ujhelyi_Gabriella_2012.pdf 1 4D021A0F9F4988B60F8919167796781C 9726de87b297f81a947be3506c3c605e1be5d5e4f174bc618d64f16f8c82253c 802039df-52de-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27898 Nacionalni inštitut za biologijo 0 0 0