TITLE:
Development of a Multiplex Ligation Dependent Probe Amplification (MLPA) Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events
AUTHORS:
Patrick Guertler, Ottmar Goerlich, Ulrich Busch
KEYWORDS:
MLPA, Genetically Modified Organism, Rapeseed, Capillary Electrophoresis, Detection
JOURNAL NAME:
Agricultural Sciences,
Vol.5 No.6,
May
23,
2014
ABSTRACT:
For the official surveillance of genetically modified crops, efficient
and simple detection methods need to be on hand to implement the strict
requirements regarding approval, labelling and traceability determined by the
European Union. Therefore, a multiplex ligation dependent probe amplification
(MLPA) module was developed for simultaneous detection of five genetically
modified rapeseed events (MS8, RF3, GT73, Falcon GS40/90 and T45). Probes were
designed and concentrations were adapted in order to obtain high sensitivity
and specificity. This MLPA module was validated using certified reference
materials and its applicability was tested analyzing routine honey, mustard and
rapeseed samples. The limit of detection was determined by analyzing a dilution
series (n = 16 for each concentration) of respective transgenic DNA. After
optimization, the MLPA revealed limits of detection between 10 to 50 copies of
the transgene DNA/assay. The method proved to be sensitive and highly
reproducible. When analyzing routine samples, results obtained applying the
MLPA module were similar compared to real-time PCR.