R-Biopharm RIDAGENE CMV PG9005 Bedienungsanleitung Seite 26

3. Test principle
®
RIDA GENE CMV is a real-time PCR for the direct qualitative and quantitative
detection of cytomegalovirus in human plasma (EDTA) specimens (quantitative) and
urine (qualitative). After DNA isolation, the specific gene fragment for CMV (MIEA, if
present) is amplified.
The amplified target sequences are detected using minor groove binder (MGB)
probes attached at one end to the quencher and at the other end to a reporter
fluorescent dye (fluorophore). The probes are activated when they hybridize with the
target sequence. This bond is additionally stabilized by the binding of the MGB in the
minor groove of the dsDNA. At this point, the reporter emits a fluorescent signal that
is detected by the optical unit of real-time PCR device. The fluorescent signal
increases with the quantity of amplicons formed. For quantification, the signal is
compared with the standard curves running in parallel and output in genome
equivalents/reaction (gEq/reaction). A factor (gEq/ml) that takes into account the
dilutions of the DNA extraction (depending on the extraction device used) and the
PCR approach are used to determine the DNA content of the specimen in genome
equivalents/ml (gEq/ml). Conversion to international units/ml (IU/ml) is done using a
correction factor (IU/ml) that also takes into account the traceability to the 1
International Standard for Human Cytomegalovirus.
®
The RIDA GENE CMV test contains an Internal Control DNA (ICD) to control the
specimen preparation and potential PCR inhibition.
4. Reagents provided
Table 1: Reagents provided (The reagents in a kit are sufficient for 100 determinations.)
Kit code Reagents
1
Master Mix
D
Internal Control DNA
P
Positive Control
10^2
Standard A
10^3
Standard B
10^4
Standard C
10^5
Standard D
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3
Quantity
4x 540 µl
2x 600 µl
2x 220 µl
1x 220 µl
1x 220 µl
1x 220 µl
1x 220 µl
2019-05-28
st
WHO
Lid color
yellow
black
blue
violet
violet
violet
violet

RIDA GENE CMV