Plasmid AmpC: CIT / DHA PCR Kit CE/IVD
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RealCycler CIDH-
Catalog Number: RealCycler CIDH-
1 Intended use:
RealCycler AMPC is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of plasmid mediated ampC: ACC, EBC, CIT and DHA DNA in clinical samples.
The system includesan internal control CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition.
2 Principle of the test
The polymerase chain reaction (PCR) is based on the amplification of a specific region of the DNA/RNA by using complementary primers to the target sequence. Real-time PCR uses marked probes with fluorophores that emit fluorescence in the case of amplification. The cycle of the PCR protocol in which appears significant fluorescence is proportional to the DNA/RNA quantity present in the sample. This value is called Cycle Threshold (Ct) or Cycle Quantification (Cq).
In the AMPC1 amplification, ACC is detected in the corresponding channel of FAM fluorophore,CHIC is detected in the corresponding channel of Alx532 fluorophore andEBC is detected in the corresponding channel of TxRfluorophore. In the AMPC2 amplification, CIT is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 fluorophore and DHA is detected in the corresponding channel of TxRfluorophore.
3 Technical specifications
Sensitivity
- ACC: 100 copies/μL.
- EBC: 500 copies/μL.
- CIT:10 copies/μL.
- DHA:1 copy/μL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity
- ACC: ACC gene.
- EBC: ACT gene.
- CIT: CMY gene.
- DHA: DHA gene.
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler AMPC includes the AMPC1 AmpliMix and AMPC2 AmpliMix dispensed into amplification tubes, an ACC-EBC DNA Positive Control, which contains a mixture of ACC and EBC DNA and a CIT-DHA DNA Positive Control,whichcontainsa mixture of CIT and DHA DNA.
All reagents are ready to use without adding or rebuilding any component.
RealCycler AMPC is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of plasmid mediated ampC: ACC, EBC, CIT and DHA DNA in clinical samples.
The system includesan internal control CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition.
2 Principle of the test
The polymerase chain reaction (PCR) is based on the amplification of a specific region of the DNA/RNA by using complementary primers to the target sequence. Real-time PCR uses marked probes with fluorophores that emit fluorescence in the case of amplification. The cycle of the PCR protocol in which appears significant fluorescence is proportional to the DNA/RNA quantity present in the sample. This value is called Cycle Threshold (Ct) or Cycle Quantification (Cq).
In the AMPC1 amplification, ACC is detected in the corresponding channel of FAM fluorophore,CHIC is detected in the corresponding channel of Alx532 fluorophore andEBC is detected in the corresponding channel of TxRfluorophore. In the AMPC2 amplification, CIT is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 fluorophore and DHA is detected in the corresponding channel of TxRfluorophore.
3 Technical specifications
Sensitivity
- ACC: 100 copies/μL.
- EBC: 500 copies/μL.
- CIT:10 copies/μL.
- DHA:1 copy/μL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity
- ACC: ACC gene.
- EBC: ACT gene.
- CIT: CMY gene.
- DHA: DHA gene.
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler AMPC includes the AMPC1 AmpliMix and AMPC2 AmpliMix dispensed into amplification tubes, an ACC-EBC DNA Positive Control, which contains a mixture of ACC and EBC DNA and a CIT-DHA DNA Positive Control,whichcontainsa mixture of CIT and DHA DNA.
All reagents are ready to use without adding or rebuilding any component.
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