An innovative test for non-invasive Kell genotyping on circulating fetal DNA by means of the allelic discrimination of K1 and K2 antigens.
- Resource Type
- Article
- Authors
- Cro', Fabiana; Lapucci, Cristina; Vicari, Emilio; Salsi, Ginevra; Rizzo, Nicola; Farina, Antonio
- Source
- American Journal of Reproductive Immunology. Dec2016, Vol. 76 Issue 6, p499-503. 5p.
- Subject
- *KELL blood group system
*ANTIGENS
*DNA
*POLYMERASE chain reaction
*FETUS
- Language
- ISSN
- 1046-7408
Objective The aim of this study was to present a new method for fetal Kell genotyping by means of the allelic discrimination of K1 and K2 in real-time polymerase chain reaction ( PCR). Methods Real-time quantitative polymerase chain reaction incorporating an allele-specific primer was developed for detecting the K allele of KEL. Results By means of this method, the K1/K2 genotype was able to be determined in all blood samples analyzed. Results using cell-free fetal DNA (cff DNA) from two Kell-negative pregnant women confirmed the Kell-positive genotype of fetuses. The real-time PCR analysis also allowed the determination of the fetal fraction using the quantification of Kell-positive DNA. Conclusion An efficient and reliable strategy for Kell genotyping is herein presented. The method was optimized on cff DNA to create a non-invasive prenatal test which could be routinely used for the prevention of hemolytic disease of the fetus and the newborn ( HDFN). [ABSTRACT FROM AUTHOR]