Hong Yun Ji, Kim Hyungsuk, Song Eun Young, Cho Duck, Park Kyoung Un, Han Kyou-Sup
Clin Lab. 2016 Oct 1;62(10):1995-2000. doi: 10.7754/Clin.Lab.2016.160320.
DEL, a variant of RhD, is difficult to detect in routine blood bank testing owing to its extremely low levels of D antigen expression. However, DEL is capable of alloimmunizing a patient when transfused into RhDnegative individuals.
In this study, we developed real-time PCR and melting curve analysis for the rapid detection of the DEL phenotype.
Of the 325 serologically RhD-negative individuals involved in the study, 56 (17.2%) had melting temperatures distinguishable from complete RHD absence as follows: 53 RHD (c.1227G>A) DEL specimens had a plateau at 54 - 56°C and a peak at 61.95°C, while 3 RHD (c.1222T>C) DEL had a melting temperature of 62.62°C. All DEL results were identical to those obtained by multiplex single-base primer extension reactions.
The rapid DEL genotyping method developed in this study will be useful for screening DEL in serologically RhD-negative donors and preventing the alloimmunization of RhD-negative individuals by DEL.
DEL是RhD的一种变体，由于其D抗原表达水平极低，在常规血库检测中难以检测到。然而，DEL输入RhD阴性个体时能够使患者产生同种免疫。
在本研究中，我们开发了实时PCR和熔解曲线分析方法用于快速检测DEL表型。
参与本研究的325名血清学检测为RhD阴性的个体中，56名（17.2%）的熔解温度与完全不存在RHD的情况可区分，如下：53份RHD（c.1227G>A）DEL标本在54 - 56°C有一个平台期，在61.95°C有一个峰值，而3份RHD（c.1222T>C）DEL的熔解温度为62.62°C。所有DEL结果与多重单碱基引物延伸反应获得的结果一致。
本研究开发的快速DEL基因分型方法将有助于在血清学检测为RhD阴性的献血者中筛查DEL，并防止DEL导致RhD阴性个体发生同种免疫。
