摘要
目的无创产前检测胎儿非整倍体(NIPT)自制血浆质控物的研制及其性能验证。方法对2019年1月1日到2021年6月30日来绍兴市妇幼保健院遗传咨询门诊就诊、并选择NIPT检测结果为阴性的孕妇随访新生儿性别,将对应储存的孕妇血浆分成男婴孕妇组19份、女婴孕妇组120份。顺序收集2020年9月至2021年9月的9360例病例作为临床验证病例。第1步:用1例47岁未孕健康女性空白血浆作为基质,每管200μl血浆分别加入5个不同体积(0.1、0.2、0.5、2.5和5μl)的胎儿染色体非整倍体(T21、T18、T13)检测试剂盒自带的阳性DNA,根据检测结果选择0.5μl的阳性DNA加样量。第2步:将男婴孕妇组血浆(19份)和女婴孕妇组血浆(20份)分别作为基质,每管(205μl)加入0.5μl的阳性DNA,发现女婴孕妇组血浆的检测结果重复性好,敏感度为100%。第3步:对120份女婴孕妇组血浆的自制血浆质控品进行性能评价,通过计算Z值、胎儿游离DNA浓度的CV值变化等指标来评价储存稳定性、基质均匀性、加入不同批号阳性DNA时的影响。结果女婴孕妇组混合血浆为基质重复性好,敏感度为100%,而男婴孕妇组留存血浆为基质的敏感度只有84%。批内重复性结果显示女婴孕妇组血浆作为基质中胎儿游离DNA浓度的CV值为3.9%;加入0.5μl阳性DNA后,自制阳性血浆质控品的胎儿游离DNA浓度为5.63%±0.42%,Z值均>6,放置3个月,胎儿游离DNA浓度的CV值为7%。阳性DNA的批号不同,浓度会有差异,当阳性DNA的胎儿游离DNA浓度值低于10%时,要加入1μl的阳性DNA。经2020年9月至2021年9月9360例临床检测中的应用,发现所有阳性血浆质控物均呈阳性结果,患者21三体的阳性预测值为100%。结论成功研制NIPT自制阳性血浆质控物,初步实验结果证实重复性好,较为稳定,适合在临床上推广。
Objective To develop a self-made plasma quality control material for non-invasive prenatal testing(NIPT)and evaluate its performance.Methods 139 NIPT-negative maternal plasmas stored in the genetic department of Shaoxing maternal and child health hospital from January 1,2019 to June 30,2021 were divided into male groups(19 cases)and female groups(120 cases)according to the neonatal gender.9360 cases from September 2020 to September 2021 were enrolled as clinical validation cases.First step,200μl plasma from a 47 years-old non-pregnant healthy women was used as a matrix.Different amounts(0.1,0.2,0.5,2.5,and 5μl)of positive DNA from fetal chromosome aneuploidy(T21,T18,T13)detection kit were added.The appropriate volume of positive DNA was 0.5μl according to the test results.Second step,Plasma in male and female group was treated as matrix.0.5μl positive DNA was added per 205μl.Plasma matrix from female group showed good repeatability and the sensitivity was 100%.Third step,evaluate the self-made plasma quality control material,including storage stability,matrix uniformity and repeatability,and the effect of different batch numbers of positive DNA,by calculating Z score and the CV of fetal DNA concentration(FF).Results Plasma matrix from female group showed good repeatability and the sensitivity was 100%,while the sensitivity of male group was only 84%.The CV of FF in female matrix was 3.9%in the repetitive experiments.After adding 0.5μl positive DNA,the mean FF of self-made positive plasma quality control was 5.63%±0.42%,Z values>6,and the CV was 7%after storage of three months.Considering the concentration variation of positive DNA in different lots,1μl of positive DNA should be added when the FF of positive DNA is lower than 10%.Used in 9360 clinical cases from September 2020 to September 2021,all positive plasma quality control materials showed positive results,and the positive predictive value of trisomy 21 was 100%.Conclusions The NIPT self-made positive plasma quality control material has been success
作者
曾艳
张涛
钱燕娣
高玉玲
熊程
倪蕴勤
罗婷婷
张丽芳
钱飞燕
Zeng Yan;Zhang Tao;Qian Yandi;Gao Yuling;Xiong Cheng;Ni Yunqin;Luo Tingting;Zhang Lifang;Qian Feiyan(Genetic department of Shaoxing Maternity and Child Health Care Hospital,Shaoxing 312000,China)
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2022年第5期444-448,共5页
Chinese Journal of Laboratory Medicine
基金
绍兴市妇幼保健院科技研究发展基金(2021YF001)。
关键词
新生儿筛查
血浆质控品
无创产前筛查
胎儿非整倍体
胎儿游离DNA
Neonatal screening
Plasma quality control product
Non-invasive prenatal testing
Aneuploidy
Cell-free fetal DNA
