A High-Throughput Multiplex Method Adapted for GMO Detection

Abstract

A high-throughput multiplex assay for the detection of genetically modified organisms (GMO) was developed on the basis of the existing SNPlex method designed for SNP genotyping. This SNPlex assay allows the simultaneous detection of up to 48 short DNA sequences (approximately 70 bp; "signature sequences") from taxa endogenous reference genes, from GMO constructions, screening targets, construct-specific, and event-specific targets, and finally from donor organisms. This assay avoids certain shortcomings of multiplex PCR-based methods already in widespread use for GMO detection. The assay demonstrated high specificity and sensitivity. The results suggest that this assay is reliable, flexible, and cost- and time-effective for high-throughput GMO detection.

Four new SYBR®Green qPCR screening methods for the detection of Roundup Ready®, LibertyLink®, and CryIAb traits in genetically modified products

“…Several multiplex qPCR formats are available reducing the number of analyses and facilitating high throughput but requiring multi-channel detection devices and often including costly detection probes for at least some of the targets [8][9][10]. In other cases, the applied chemistries are quite complex or are to be combined with other technologies that are less appropriate for routine applications [11][12][13][14][15][16]. Several of these approaches involve the use of PCR with multiple targets and consecutive detection and identification of the amplification products using microarrays [11][12][13].…”

“…In other cases, the applied chemistries are quite complex or are to be combined with other technologies that are less appropriate for routine applications [11][12][13][14][15][16]. Several of these approaches involve the use of PCR with multiple targets and consecutive detection and identification of the amplification products using microarrays [11][12][13]. Apart from requiring additional costly array analysis equipment, these approaches are often prone to variable quality of the array chips making them less suitable for routine or enforcement purposes [17].…”

Development of a Molecular Platform for GMO Detection in Food and Feed on the Basis of “Combinatory qPCR” Technology

“…At the present time several high-tech strategies like multiplex PCR and consecutive detection and identification of the amplification products using micro-arrays (Chaouachi et al, 2008, Morisset et al, 2008, Hamels et al, 2009 or PCR combined with capillary electrophoresis (Nadal et al, 2009) have been proposed to deal with this discriminative problem and the broad diversity of GMO. However, at the present time, these technologies require additional costly equipment and investments in technical support.…”

Reliability of PCR based screening for identification and quantification of GMOs

“…For that reason, laboratories call for simple cost effective assays as suggested e.g. by Hamels et al (2009) or Chaouachi et al (2008. However, they need to be approved by practice.…”