一种通过K1和K2抗原的等位基因鉴别对循环胎儿DNA进行无创Kell基因分型的创新检测方法。
An innovative test for non-invasive Kell genotyping on circulating fetal DNA by means of the allelic discrimination of K1 and K2 antigens.
作者信息
Cro' Fabiana, Lapucci Cristina, Vicari Emilio, Salsi Ginevra, Rizzo Nicola, Farina Antonio
机构信息
Geneticlab, Noventa Vicentina, Italy.
Division of Prenatal Medicine, Department of Medicine and Surgery (DIMEC), University of Bologna, Bologna, Italy.
出版信息
Am J Reprod Immunol. 2016 Dec;76(6):499-503. doi: 10.1111/aji.12593. Epub 2016 Oct 11.
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 (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.
CONCLUSION
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).
目的
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。
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。
结果
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。
结论
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。
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