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An innovative test for non‐invasive Kell genotyping on circulating fetal DNA by means of the allelic discrimination of K1 and K2 antigens
Author(s) -
Cro' Fabiana,
Lapucci Cristina,
Vicari Emilio,
Salsi Ginevra,
Rizzo Nicola,
Farina Antonio
Publication year - 2016
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/aji.12593
Subject(s) - genotyping , genotype , polymerase chain reaction , primer (cosmetics) , allele , fetus , cell free fetal dna , biology , real time polymerase chain reaction , dna , microbiology and biotechnology , immunology , prenatal diagnosis , genetics , pregnancy , gene , chemistry , organic chemistry
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 ).