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Published on: February 20, 2012
A new QRT-PCR assay designed for the differentiation between elements provided from Agrobacterium sp. in GMOs plant
Nesrine Nabi1, Maher Chaouachi1, Mohamed Salem Zellama1
1Laboratoire de Génétique Biodiversité et Valorisation des Bio-ressources, Institut Supérieur de Biotechnologie de Monastir (ISBM), Université de Monastir, Avenue Tahar Haddad, 5000, BP74, Monastir, Tunisia.
Abstract:
The question asked in the present work was how to differentiate between contamination of field samples with and GM plants contained sequences provided from this bacterium in order to avoid false positives in the frame of the detection and the quantification of GMO. For this, new set of primers and corresponding TaqMan Minor Groove Binder (MGB) probes were designed to target Agrobacterium sp. using the tumor-morphology-shooty gene (TMS1). Final standard curves were calculated for each pathogen by plotting the threshold cycle value against the bacterial number (log (colony forming units) per milliliter) via linear regression. The method designed was highly specific and sensitive, with a detection limit of 10CFU/ml. No significant cross-reaction was observed. Results from this study showed that TaqMan real-time PCR, is potentially an effective method for the rapid and reliable quantification of Agrobacterium sp. in samples containing GMO or non GMO samples.
