blaIMP + blaNDM-1 / blaOXA + blaVIM + blaKPC + blaCTX-M PCR Kit CE/IVD
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RealCycler INOCVK-
Catalog Number: RealCycler INOCVK-
1 Intended use:
RealCycler INOCVK is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of beta-lactamaseblaIMPgene, blaNDM-1 gene (New Delhi metallo-beta-lactamase), carbapenemases blaOXA gene, beta-lactamase blaCTX-M gene, metallobetalactamases blaVIM gene and carbapenemases blaKPC genes in clinical samples.
The system includes an internal control CHIC (Competitive Heterologous Internal Control) to prevent false negativesdueto 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 fluorophoresthat 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 IMND amplification, blaIMP gene is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore and blaNDM-1 geneis detected in the corresponding channel of TxR fluorophore. In the OCTVIK amplification, blaOXAgeneis detected in the corresponding channel of FAM fluorophore, blaCTX-M gene is detected in the corresponding channel of Alx532 or HEX fluorophore, blaVIMgene is detected in the corresponding channel of TxR fluorophore and blaKPCgene is detected in the corresponding channel of Alx647 or Cy5 fluorophore.
3 Technical specifications
Sensitivity
- blaIMP: 10 copies/μL.
-blaNDM-1: 100 copies/μL.
-blaOXA-48: 10 copies/μL.
-blaCTX-M1: 1 copy/μL.
-blaCTX-M2: 10 copies/μL.
-blaCTX-M9: 1 copy/μL.
-blaVIM: 10 copies/μL.
-blaKPC: 10 copies/μL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity
- blaIMP: blaIMPgene.
- blaNDM-1: blaNDM-1 gene.
- blaOXA: blaOXAgene (blaOXA-48, blaOXA-162, blaOXA-163, blaOXA-244, blaOXA-245, blaOXA-247, blaOXA-370 genes and eventually, other sequences of blaOXAgroup).
- blaCTX-M: blaCTX-M gene.
- blaVIM: blaVIMgene.
- blaKPC: blaKPC gene.
4 Contents
RealCycler INOCVK-U / INOCVK-G includes the IMND AmpliMix and OCTVIK AmpliMix, an IMND DNA Positive Control, which contains a mixture of blaIMPand blaNDM-1 DNA, and an OCTVIK DNA Positive Control which contains a mixture ofblaOXA-48, blaCTX-M2, blaCTX-M9, blaVIMand blaKPC DNA. Each sample should be simultaneously analysedwith both AmpliMix (IMND and OCTVIK). OCTVIK AmpliMix cannot be used individually given that CHIC is not included.
All reagents are ready to use without adding or rebuilding any component.
RealCycler INOCVK is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of beta-lactamaseblaIMPgene, blaNDM-1 gene (New Delhi metallo-beta-lactamase), carbapenemases blaOXA gene, beta-lactamase blaCTX-M gene, metallobetalactamases blaVIM gene and carbapenemases blaKPC genes in clinical samples.
The system includes an internal control CHIC (Competitive Heterologous Internal Control) to prevent false negativesdueto 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 fluorophoresthat 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 IMND amplification, blaIMP gene is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore and blaNDM-1 geneis detected in the corresponding channel of TxR fluorophore. In the OCTVIK amplification, blaOXAgeneis detected in the corresponding channel of FAM fluorophore, blaCTX-M gene is detected in the corresponding channel of Alx532 or HEX fluorophore, blaVIMgene is detected in the corresponding channel of TxR fluorophore and blaKPCgene is detected in the corresponding channel of Alx647 or Cy5 fluorophore.
3 Technical specifications
Sensitivity
- blaIMP: 10 copies/μL.
-blaNDM-1: 100 copies/μL.
-blaOXA-48: 10 copies/μL.
-blaCTX-M1: 1 copy/μL.
-blaCTX-M2: 10 copies/μL.
-blaCTX-M9: 1 copy/μL.
-blaVIM: 10 copies/μL.
-blaKPC: 10 copies/μL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity
- blaIMP: blaIMPgene.
- blaNDM-1: blaNDM-1 gene.
- blaOXA: blaOXAgene (blaOXA-48, blaOXA-162, blaOXA-163, blaOXA-244, blaOXA-245, blaOXA-247, blaOXA-370 genes and eventually, other sequences of blaOXAgroup).
- blaCTX-M: blaCTX-M gene.
- blaVIM: blaVIMgene.
- blaKPC: blaKPC gene.
4 Contents
RealCycler INOCVK-U / INOCVK-G includes the IMND AmpliMix and OCTVIK AmpliMix, an IMND DNA Positive Control, which contains a mixture of blaIMPand blaNDM-1 DNA, and an OCTVIK DNA Positive Control which contains a mixture ofblaOXA-48, blaCTX-M2, blaCTX-M9, blaVIMand blaKPC DNA. Each sample should be simultaneously analysedwith both AmpliMix (IMND and OCTVIK). OCTVIK AmpliMix cannot be used individually given that CHIC is not included.
All reagents are ready to use without adding or rebuilding any component.
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