Detection method details
Name: |
QL-ELE-cat-F/R |
Description: |
Qualitative real-time PCR (TaqMan) method for detection of the chloramphenicol marker potentially harboured by GMM in food enzyme preparations (Turgeon et al. 2008).
In term of food and feed safety, to evaluate the level of risks of likely AMR (antimicrobial resistance) gene acquisition by pathogens and gut microbiota, it is essential to determine if the full-length AMR genes are present. In order to assess the presence of the full-length cat gene a successive nested-PCR method can be performed to amplify a large fragment of the cat gene be performed (Fraiture et al. 2020) (see Related Methods QL-ELE-cat-F1/R1 and QL-ELE-cat-F2/R2). |
Comment: |
Target is the chloramphenicol acetyl-transferase (cat) gene conferring resistance to chloramphenicol (CmR). |
Validation: |
unknown |
Standardisation: |
none |
Related Methods: |
QL-ELE-cat-F1/R1, QL-ELE-cat-F2/cat-R2 |
Type: |
element-specific |
Target DNA element: |
V-chloramphenicol acetyl transferase
|
-
Oligonucleotides:
Forward Primer
Name:
cat-FSequence:
GTGACAAGGGTGATAAACTCAAATACSize:
26
Reverse Primer
Name:
cat-RSequence:
TGTATAAAGTGGCTCTAACTTATCCCSize:
26
Probe
Name:
cat-PSequence:
ACCTAACTCTCCGTCGCTATTGTAACCAGTSize:
30
-
Amplicon:
Sequence:
GTGACAAGGGTGATAAACTCAAATACAGCTTTTAGAACTGGTTACAATAGCGACGGAGAGTTAGGTTATTGGGATAAGTTAGAGCCACTTTATACASize:
96