Mycoplasma hominis + Ureaplasma parvum / Mycoplasma genitalium + Trichomonas vaginalis + Ureaplasma urealyticum PCR Kit CE/IVD
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RealCycler GENI-
Catalog Number: RealCycler GENI-
1 Intended use:
RealCycler GENI-U / GENI-G is a kit of reagents which allows real-time PCR detection of Mycoplasma hominis, Ureaplasma parvum, Mycoplasma genitalium, Trichomonas vaginalis and Ureaplasma urealyticum DNA in clinical samples.
Ureaplasma urealyticumpathogen could be quantified if this product is used in combination with the corresponding calibrator (reference RealCycler UREQ).
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).
In the MHUP amplification, Mycoplasma hominis is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore and Ureaplasma parvum is detected in the corresponding channel of Alx647 orCy5 fluorophore.
In the MGTVUU amplification, Mycoplasma genitalium is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore, Trichomonas vaginalis is detected in the corresponding channel of TxR fluorophore and Ureaplasma urealyticum is detected in the corresponding channel of Alx647 or Cy5 fluorophore.
3 Technical specifications
Sensitivity
Mycoplasma hominis: 50 copies/μL.
Ureaplasma parvum: 1 copy/μL.
- Mycoplasma genitalium: 100 copies/μL.
- Trichomonas vaginalis: 10 copies/μL.
- Ureaplasmaurealyticum: 100 copies/μL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity:
Mycoplasma hominis: Glyceraldehyde-3-P-dehydrogenase (gap).
Ureaplasmaparvum: ureasegene.
Mycoplasma genitalium: adhesin gene (mgpB).
Trichomonas vaginalis: 2-kb repeated sequence.
Ureaplasma urealyticum: urease gene.
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler GENI-U / GENI-G includes the MHUP AmpliMix andMGTVUU AmpliMix, a MHUP DNA Positive Control, which contains a mixture of Mycoplasma hominis and Ureaplasma parvum DNA and a MGTVUU DNA Positive Control, which contains a mixture of Mycoplasma genitalium, Trichomonas vaginalis and Ureaplasma urealyticum.
RealCycler GENI-U / GENI-G is a kit of reagents which allows real-time PCR detection of Mycoplasma hominis, Ureaplasma parvum, Mycoplasma genitalium, Trichomonas vaginalis and Ureaplasma urealyticum DNA in clinical samples.
Ureaplasma urealyticumpathogen could be quantified if this product is used in combination with the corresponding calibrator (reference RealCycler UREQ).
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).
In the MHUP amplification, Mycoplasma hominis is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore and Ureaplasma parvum is detected in the corresponding channel of Alx647 orCy5 fluorophore.
In the MGTVUU amplification, Mycoplasma genitalium is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 or HEX fluorophore, Trichomonas vaginalis is detected in the corresponding channel of TxR fluorophore and Ureaplasma urealyticum is detected in the corresponding channel of Alx647 or Cy5 fluorophore.
3 Technical specifications
Sensitivity
Mycoplasma hominis: 50 copies/μL.
Ureaplasma parvum: 1 copy/μL.
- Mycoplasma genitalium: 100 copies/μL.
- Trichomonas vaginalis: 10 copies/μL.
- Ureaplasmaurealyticum: 100 copies/μL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity:
Mycoplasma hominis: Glyceraldehyde-3-P-dehydrogenase (gap).
Ureaplasmaparvum: ureasegene.
Mycoplasma genitalium: adhesin gene (mgpB).
Trichomonas vaginalis: 2-kb repeated sequence.
Ureaplasma urealyticum: urease gene.
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler GENI-U / GENI-G includes the MHUP AmpliMix andMGTVUU AmpliMix, a MHUP DNA Positive Control, which contains a mixture of Mycoplasma hominis and Ureaplasma parvum DNA and a MGTVUU DNA Positive Control, which contains a mixture of Mycoplasma genitalium, Trichomonas vaginalis and Ureaplasma urealyticum.
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