title: An innovative test for non-invasive Kell genotyping on circulating fetal DNA by means of the allelic discrimination of K1 and K2 antigens.
abstract: 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).
  
  
  
  Real-time quantitative polymerase chain reaction incorporating an allele-specific primer was developed for detecting the K allele of KEL.
  
  
  
  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 (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.
  
  
  
  An 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).
authors.affiliation: Geneticlab, Noventa Vicentina, Italy.
authors.forename: Fabiana
authors.initials: F
authors.lastname: Cro'
authors.affiliation: Geneticlab, Noventa Vicentina, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Lapucci
authors.affiliation: Geneticlab, Noventa Vicentina, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Vicari
authors.affiliation: Division of Prenatal Medicine, Department of Medicine and Surgery (DIMEC), University of Bologna, Bologna, Italy.
authors.forename: Ginevra
authors.initials: G
authors.lastname: Salsi
authors.affiliation: Division of Prenatal Medicine, Department of Medicine and Surgery (DIMEC), University of Bologna, Bologna, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Rizzo
authors.affiliation: Division of Prenatal Medicine, Department of Medicine and Surgery (DIMEC), University of Bologna, Bologna, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Farina
mesh_terms: D000328:Adult; D000483:Alleles; D000073888:Cell-Free Nucleic Acids / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D017931:DNA Primers / Q000737:chemistry / Q000378:metabolism; D004899:Erythroblastosis, Fetal / Q000097:blood / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D005260:Female; D005333:Fetus; D015870:Gene Expression; D005838:Genotype; D060005:Genotyping Techniques; D006801:Humans; D007231:Infant, Newborn; D007626:Kell Blood-Group System / Q000235:genetics* / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology; D008666:Metalloendopeptidases / Q000235:genetics* / Q000276:immunology; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D000073888:Cell-Free Nucleic Acids; D017931:DNA Primers; D007626:Kell Blood-Group System; D008562:Membrane Glycoproteins; C585978:KEL protein, human; D008666:Metalloendopeptidases
keywords: circulating free fetal DNA; kell genotype; real-time PCR