Cro' Fabiana, Lapucci Cristina, Vicari Emilio, Salsi Ginevra, Rizzo Nicola, Farina Antonio
Geneticlab, Noventa Vicentina, Italy.
Division of Prenatal Medicine, Department of Medicine and Surgery (DIMEC), University of Bologna, Bologna, Italy.
Am J Reprod Immunol. 2016 Dec;76(6):499-503. doi: 10.1111/aji.12593. Epub 2016 Oct 11.
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).
本研究的目的是通过实时聚合酶链反应（PCR）中K1和K2的等位基因鉴别，提出一种新的胎儿Kell基因分型方法。
开发了一种结合等位基因特异性引物的实时定量聚合酶链反应，用于检测KEL的K等位基因。
通过这种方法，能够在所有分析的血样中确定K1/K2基因型。使用两名Kell阴性孕妇的游离胎儿DNA（cffDNA）的结果证实了胎儿的Kell阳性基因型。实时PCR分析还可以通过定量Kell阳性DNA来确定胎儿分数。
本文提出了一种高效可靠的Kell基因分型策略。该方法在cffDNA上进行了优化，以创建一种可常规用于预防胎儿和新生儿溶血病（HDFN）的非侵入性产前检测方法。
