Plasmid AmpC-type β-lactamases: blaCIT / blaDHA / blaACC and blaEBC PCR Kit CE/IVD
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RealCycler CIDHAE-
Catalog Number: RealCycler CIDHAE-
1 Intended use:
RealCycler CIDHAE-U / CIDHAE-G is an in vitro diagnostic kit of reagents which allows real-time PCR detection of plasmid AmpC-type β-lactamases DNA: blaCIT, blaDHA, blaACC and blaEBC simultaneously in clinical samples.
The system includes an 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).
he amplification of blaCIT is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore, blaDHA is detected in the corresponding channel of TxR fluorophore and blaACC and blaEBC is detected in the corresponding channel of Alx647 or Cy5 fluorophore.
3 Technical specifications
Sensitivity
- blaCIT: 1 copy/μL.
- blaDHA: 5 copies/μl.
- blaACC: 1 copy/μl.
- blaEBC: 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
- blaCIT: laCMYgene (blaCMY-2, 4, 7,14, 15, 17, 18, 20, 21, 22, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 38, 39, 40, 42, 43, 44, 46, 47, 49, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 64, 66, 68, 69, 71, 72, 76, 77, 78, 79, 80, 81, 84, 90, 102, 104, 110 and 111; it is notexcluded the detection of other sequences of blaCMY group).
- blaDHA: blaDHA gene (blaDHA-1, 3, 5, 6 and 7; it is not excluded the detection of other sequences of blaDHA group).
- blaACC: blaACC gene (blaACC-1, 4, 5 and 6; it is not excluded thedetection of other sequences of blaACC group).
- blaEBC: blaACT gene (blaACT-2, 3, 8, 10, 13, 20, 21 and 34; it is notexcluded the detection of other sequences of blaACT group). blaMIR gene (blaMIR-1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15; it is not excluded thedetectionof other sequences of blaMIR group).
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler CIDHAE-U / CIDHAE-G includes the CIDHAE AmpliMix and a CIDHAE DNA Positive Control, which contains a mixture of blaCIT (blaCMY), blaDHA, blaACC and blaEBC (blaACT and blaMIR) DNA.
All reagents are ready to use without adding or rebuilding any component.
RealCycler CIDHAE-U / CIDHAE-G is an in vitro diagnostic kit of reagents which allows real-time PCR detection of plasmid AmpC-type β-lactamases DNA: blaCIT, blaDHA, blaACC and blaEBC simultaneously in clinical samples.
The system includes an 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).
he amplification of blaCIT is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore, blaDHA is detected in the corresponding channel of TxR fluorophore and blaACC and blaEBC is detected in the corresponding channel of Alx647 or Cy5 fluorophore.
3 Technical specifications
Sensitivity
- blaCIT: 1 copy/μL.
- blaDHA: 5 copies/μl.
- blaACC: 1 copy/μl.
- blaEBC: 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
- blaCIT: laCMYgene (blaCMY-2, 4, 7,14, 15, 17, 18, 20, 21, 22, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 38, 39, 40, 42, 43, 44, 46, 47, 49, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 64, 66, 68, 69, 71, 72, 76, 77, 78, 79, 80, 81, 84, 90, 102, 104, 110 and 111; it is notexcluded the detection of other sequences of blaCMY group).
- blaDHA: blaDHA gene (blaDHA-1, 3, 5, 6 and 7; it is not excluded the detection of other sequences of blaDHA group).
- blaACC: blaACC gene (blaACC-1, 4, 5 and 6; it is not excluded thedetection of other sequences of blaACC group).
- blaEBC: blaACT gene (blaACT-2, 3, 8, 10, 13, 20, 21 and 34; it is notexcluded the detection of other sequences of blaACT group). blaMIR gene (blaMIR-1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15; it is not excluded thedetectionof other sequences of blaMIR group).
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler CIDHAE-U / CIDHAE-G includes the CIDHAE AmpliMix and a CIDHAE DNA Positive Control, which contains a mixture of blaCIT (blaCMY), blaDHA, blaACC and blaEBC (blaACT and blaMIR) DNA.
All reagents are ready to use without adding or rebuilding any component.
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