An innovative test for non-invasive Kell genotyping on circulating fetal DNA by means of the allelic discrimination of K1 and K2 antigens

作者：Cro' Fabiana; Lapucci Cristina; Vicari Emilio; Salsi Ginevra; Rizzo Nicola; Farina Antonio^*

来源：American Journal of Reproductive Immunology, 2016, 76(6): 499-503.

DOI：10.1111/aji.12593

• circulating free fetal DNA
• kell genotype
• real-time PCR

ObjectiveThe 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).
MethodsReal-time quantitative polymerase chain reaction incorporating an allele-specific primer was developed for detecting the K allele of KEL.
ResultsBy means of this method, the K1/K2 genotype was able to be determined in all blood samples analyzed. Results using cell-free fetal DNA (cffDNA) 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.
ConclusionAn efficient and reliable strategy for Kell genotyping is herein presented. The method was optimized on cffDNA 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).

• 出版日期2016-12