Description
PURPOSE
«ALSENSE RHD» Reagent kit is designed for the qualitative determination of RHD gene in peripheral blood plasma from a Rh-negative pregnant woman, which contains cell-free fetal DNA. A crucial condition that enhances the specificity of this method is its application at the earliest possible stages, from the 9th-10th week of “embryological” pregnancy in serologically Rh-negative women. The analysis employs the most effective genotyping method ‒ real-time PCR with fluorescence detection based on TaqMan technology.
ASSAY PRINCIPLE
The Reagent kit is a real-time RT-PCR-based assay with hybridization-fluorescence detection of accumulating PCR products. During the reaction, only target DNA fragments are amplified due to the primers specificity. The hybridization probe contains in its structure both a fluorescent dye and a corresponding fluorescence quencher. The hybridization probe is annealed on copies of a specific DNA site and in the next cycle of PCR is cleaved by polymerase. Cleavage of probe is accompanied by the release of the dye, and the fluorescence of the mixture increases. The fluorescence is measured by the detecting thermal cycler in real time in parallel on several channels.
BIOMATERIAL FOR ANALYSIS
- Peripheral blood plasma of a Rh-negative pregnant woman anticoagulated with EDTA-K2 or CPDA.
- Buccal epithelium samples of a woman (if necessary, for examining the mother’s DNA).
