低比例母体细胞污染对α-地中海贫血产前诊断的影响研究  

Effect of Low Proportion of Maternal Cell Contamination on Prenatal Diagnosis of α-thalassemia

作　　者：夏勇[1,2] 熊礼宽 麦光兴[1,2] XIA Yong;XIONG Likuan;MAI Guangxing(Central Laboratory,Shenzhen Baoan Women’s and Children’s Hospital,Shenzhen Guangdong 518102,China;Shenzhen Key Laboratory of Birth Defects,Shenzhen Guangdong 518102,China)

机构地区：[1]深圳市宝安区妇幼保健院中心实验室,广东深圳518102 [2]深圳市出生缺陷重点实验室,广东深圳518102

出　　处：《中国卫生标准管理》2020年第16期109-112,共4页China Health Standard Management

基　　金：2019年深圳市宝安区医疗卫生基础研究项目(2019JD378)。

摘　　要：目的研究低比例(<10%)母体细胞污染对α-地中海贫血产前诊断的影响,为后续进行相关的高灵敏度母体细胞污染鉴定提供前期基础。方法选取2019年1—10月在深圳市宝安区妇幼保健院确诊为Bart’s水肿胎儿(基因型为--SEA/--SEA)的样本4例及正常对照孕妇的样本4例(基因型为αα/αα),建立母体细胞污染浓度梯度为50%、25%、12.5%、6.25%、3.125%、1.5625%模型。研究现行常用的荧光定量PCR技术的鉴定能力及低比例母体细胞污染对α-地中海贫血基因分型结果的影响。结果荧光定量PCR技术对于母体细胞污染比例为12.5%的鉴定能力(25%,1/4)已不可靠,对于6.25%及以下均无法鉴定。同时,低至3.125%的母体细胞污染仍能够严重影响α-地中海贫血,尤其是点突变型的基因分型结果,造成误判。结论低比例母体细胞污染使用现有荧光定量PCR技术容易漏检,且影响这类α-地中海贫血基因分型结果。Objective To study the impact of low proportion(<10%)of maternal cell contamination on the prenatal diagnosis of α-thalassemia,and to provide a preliminary basis for the subsequent high sensitivity identification of cell contamination.Methods Four samples of fetuses with Bart’s dropsy(genotype--SEA/--SEA)diagnosed in Shenzhen Baoan Women’s and Children’s hospital and four samples of normal control pregnant women(genotype αα/αα)from January 2019 to October 2019 were selected to establish a model of maternal cell contamination with concentration gradient of 50%,25%,12.5%,6.25%,3.125%and 1.5625%.The identification ability of fluorescence quantitative PCR and the impact of low proportion of maternal cell contamination on the accurate genotyping ofα-thalassemia were studied.Results Fluorescent quantitative PCR was not reliable for the identification of 12.5%of the maternal cell contamination(25%,1/4),and could not be used for the identification of 6.25%or less.At the same time,as low as 3.125%of maternal cell contamination can still seriously affect the genotyping results of alpha thalassemia,especially for point mutation,resulting in misclassification.Conclusion Presently used Fluorescent quantitative PCR technology could miss the detection of low proportion of maternal cell contamination,and affects the genotyping results of this type of alpha thalassemia.

关 键 词：母体细胞污染 Α-地中海贫血 产前诊断 低比例 荧光定量PCR 基因分型 

分 类 号：R169[医药卫生—公共卫生与预防医学]