Innovative Application of Fluorescent Micro-sphere Based Assay for Multiple GMO Detection

摘要

An innovative multiple screening protocol allowing the detection of specific DNA sequences of p35S and epsps in GM soya flours simultaneously was developed. For the application of the Luminex xMAP technology, two different sets of fluorescent beads were respectively cross-linked to the specific oligonucleotide probes previously amplified and labelled by PCR reaction in the presence of a biotinylated nucleotide. The detection of the amplification products bounded by the streptavidin-phycoeritrin conjugate was performed using the Luminex-100 instrument. The system allows to identify extremely low amounts of labelled targets as of 7.8 10-4 nmol of p35S antiprobe with an associated value of RSDr equal to 2.8. Parameters such as repeatability, reproducibility, RSD, LOD and LOQ were considered to evaluate the performances of the assay. The potentiality of the system here described, enables us to look forward to a multiple target assay able to simultaneously detect and eventually quantify tens of target sequences occurring within the same sample preparation. The present study describes the first application of a multi-target fluorescent micro-sphere based assay for DNA GMO detection.