A new QRT-PCR assay designed for the differentiation between elements provided from Agrobacterium sp. in GMOs plant events and natural Agrobacterium sp . bacteria
| Autor: | Besma Mrabet, Nesrine Nabi, Harzallah Skhiri Fathia, Ahmed Ben Hafsa, Maher Chaouachi, Mohamed Salem Zellama, Khaled Said |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Detection limit Colony-forming unit Bacteria Reverse Transcriptase Polymerase Chain Reaction Agrobacterium fungi 030106 microbiology General Medicine Biology Plants Genetically Modified Real-Time Polymerase Chain Reaction biology.organism_classification Molecular biology Analytical Chemistry Microbiology Standard curve 03 medical and health sciences 030104 developmental biology Real-time polymerase chain reaction TaqMan Pathogen Food Science |
| Zdroj: | Food Chemistry. 196:58-65 |
| ISSN: | 0308-8146 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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