FR 
EN 
FetoGnost® RHD Kit allows a rapid, sensitive and non-invasive determination of the fetal Rhesus D (RhD) gene in purified samples from maternal plasma of RhD-negative pregnant women. The test is based on real-time PCR technology. Probe-specific amplification curves in VIC, FAM and NED indicate amplification of exons 5, 7 and 10 of the RHD gene in RHD-positive fetuses, respectively. In addition, an internal positive control (IPC) with detection in the Cy5 channel monitors the integrity of the kit reagents, serves as a control for DNA extraction and excludes false negative interpretation of results due to inhibition of real-time PCR. The CPI target is added during DNA extraction from maternal plasma samples.
DNA from 30 plasma samples, including controls, can be analysed in triplicate using a 96-well reaction plate (order no. HUFG100).
.
The risk of false negative results is minimised by the sensitive and robust multiplex test format of the FetoGnost® RHD Kit for the detection of three RHD gene exons in triplicate.
FetoGnost® Kit RHD has been approved for use with Applied Biosystems® 7500 instrument and QuantStudioTM 7 Pro (Thermo Fisher Scientific) (fast cycle settings not supported), but is also compatible with other real-time PCR instruments that detect and differentiate fluorescence in the FAM, VIC, NED and Cy5 channels.
When using PCR platforms not tested by ingenetix, a multiplex PCR evaluation is recommended.
Prior to any use of FetoGnost® Kit RHD, it is strongly recommended that the extraction of free fetal DNA (cff) from maternal plasma be evaluated to ensure that the extraction method produces sufficient fetal DNA. FetoGnost® Control Kit (order no. HUFG050, 50 reactions for up to 30 plasma samples) is used to assess the detection of fetal DNA.
Fragmented DNA (<300 bp) of fetal origin represents only a small proportion of the circulating free DNA (ccfDNA) present in maternal plasma and increases during pregnancy. In maternal plasma samples collected in the second trimester of pregnancy, the concentration of cffDNA can be as high as 50-200 cffDNA genomic equivalents/ml blood (Birch et al., 2005).
The official and applicable recommendations or guidelines in each country (e.g. the recommendation of DGTI, Deutsche Gesellschaft für Transfusionsmedizin und Immunhämatologie) should be followed when evaluating the FetoGnost® RHD Kit.
Sampling for the determination of fetal RHD status should not be performed before 11+0 SA.