血浆游离DNA定量检测在产前诊断中的应用效果及基因分析研究
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摘要
目的探讨对产妇进行血浆游离DNA定量检测的效果以及基因分析。方法选取我院孕晚期孕妇、唐氏高危孕妇、孕中期孕妇以及正常女性各22例,共88例,从其血浆中将游离的DNA分离出来,应用荧光PCR对SRY(仅男性含有的Y性别决定基因)、GAPDH(3-磷酸甘油醛脱氢酶基因)进行检测,统计并比较其检测的准确率、特异性、敏感度。结果孕中期孕妇与正常女性体内GAPDH的含量未出现明显差异(P>0.05),唐氏高危与孕晚期孕妇体内的GAPDH含量显著高于孕中期孕妇与正常女性(P<0.05);唐氏高危以及孕晚期孕妇的SRY含量无明显差异(P>0.05),唐氏高危与孕晚期孕妇SRY含量显著高于孕中期孕妇(P<0.05);孕晚期孕妇的检测准确率、特异性、敏感度显著高于唐氏高危与孕中期孕妇(P<0.05)。结论孕妇在产前对血浆中游离的DNA进行定量检测的效果显著,有利于产前进行无创伤的性别检测,在唐氏筛查方面具有重要意义,值得在临床上推广和应用。
Objective:To explore the effect of quantitative detection of free plasma DNA in puerpera and genetic analysis. Methods:Pregnant women late in pregnancy,Down's mid of high-risk pregnant women,pregnant women and normal women all 22 cases,a total of 88 cases,free DNA isolated from the plasma,using fluorescent PCR for SRY(men only contains the sexdetermining gene Y)and GAPDH(genes)glyceraldehyde 3-phosphoric acid dehydrogenase,statistics and compare the detection sensitivity,specificity and accuracy. Results:There was no significant difference in GAPDH between pregnant women and normal women(P>0.05). The content of GAPDH in pregnant women with high risk and late pregnancy was significantly higher than that in pregnant women(P<0.05);There was no significant difference in SRY content between the high risk and the pregnant women in late pregnancy(P>0.05). The SRY content of pregnant women with high risk and late pregnancy was significantly higher than that of pregnant women(P<0.05);The detection accuracy,specificity and sensitivity of pregnant women in late pregnancy were significantly higher than that of pregnant women with high risk and pregnancy(P<0.05). Conclusion:The pregnant women in prenatal of free DNA in plasma quantitatively detecting effect is remarkable,is advantageous to the prenatal detection of sex without trauma,in down syndrome screening is of great significance,is worth in clinical promotion and application.
Objective:To explore the effect of quantitative detection of free plasma DNA in puerpera and genetic analysis. Methods:Pregnant women late in pregnancy,Down's mid of high-risk pregnant women,pregnant women and normal women all 22 cases,a total of 88 cases,free DNA isolated from the plasma,using fluorescent PCR for SRY(men only contains the sexdetermining gene Y)and GAPDH(genes)glyceraldehyde 3-phosphoric acid dehydrogenase,statistics and compare the detection sensitivity,specificity and accuracy. Results:There was no significant difference in GAPDH between pregnant women and normal women(P>0.05). The content of GAPDH in pregnant women with high risk and late pregnancy was significantly higher than that in pregnant women(P<0.05);There was no significant difference in SRY content between the high risk and the pregnant women in late pregnancy(P>0.05). The SRY content of pregnant women with high risk and late pregnancy was significantly higher than that of pregnant women(P<0.05);The detection accuracy,specificity and sensitivity of pregnant women in late pregnancy were significantly higher than that of pregnant women with high risk and pregnancy(P<0.05). Conclusion:The pregnant women in prenatal of free DNA in plasma quantitatively detecting effect is remarkable,is advantageous to the prenatal detection of sex without trauma,in down syndrome screening is of great significance,is worth in clinical promotion and application.
引文
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[12]刘宁,赵德华,李晓乐,等.PTPS基因检测在6丙酮酰四氢蝶呤合成酶缺乏症出生后诊断和产前诊断中的应用[J].中华妇产科杂志,2016,51(12):890-894.
[13]丁圆,李东晓,刘玉鹏,等.黏多糖贮积症Ⅶ型1例患儿的临床与GUSB基因分析及其同胞的产前诊断[J].中华实用儿科临床杂志,2016,31(8):604-608.
[14]林洋洋,徐婉芳,刘彦慧,等.大规模平行测序技术在产前无创检测胎儿染色体异常中的临床应用[J].中国优生与遗传杂志,2016,1:52-54.
[15]陈梅环,黄海龙,王燕,等.东南亚缺失型遗传性持续性胎儿血红蛋白增多症伴β地中海贫血一个家系的产前诊断[J].中国实验血液学杂志,2017,25(4):1142-1146.
[2]王佳燕,陈敏,吴莉,等.无创产前基因检测双胎21、18和13-三体综合征的应用研究[J].中国实用妇科与产科杂志,2017,11(5):497-501.
[3]蔡奥捷,宗亚楠,刘宁,等.甲基丙二酸血症结合同型半胱氨酸血症家系MMACHC基因突变分析及在产前诊断中的应用[J].中华检验医学杂志,2016,39(8):613-617.
[4]Zhao X.Noninvasive prenatal testing can detect gene deletions,duplications:Use of cell-free fetal DNA for NIPT promising but not yet ready for clinical use[J].American Journal of Medical Genetics Part A,2016,170(3):556-557.
[5]邹洋,徐佩文,李杰,等.微滴式数字PCR在脊肌萎缩症基因检测和产前诊断中的应用[J].中华医学遗传学杂志,2016,33(5):594-597.
[6]郭莉,李砚.扩增孕妇血浆中游离胎儿DNA构建方法及其对产前诊断21-三体综合征中价值研究[J].中国优生与遗传杂志,2017,7(12):30-32.
[7]王红丹,冯战启,娄桂予,等.应用微阵列比较基因组杂交技术产前检测室间隔缺损胎儿2p16.3微缺失的临床研究[J].重庆医学,2016,45(23):3256-3257.
[8]Chen J,Yu C,Zhao Y,et al.A novel non-invasive detection method for the FGFR3 gene mutation in maternal plasma for a fetal achondroplasia diagnosis based on signal amplification by heminMOFs/PtNPs[J].Biosensors&Bioelectronics,2016,91(22):892-899.
[9]耿娟,刘畅,周香城,等.微滴式数字PCR技术用于检测产前诊断标本母体细胞污染的初步研究[J].中华妇产科杂志,2017,52(2):93-97.
[10]刘静,席惠,王华,等.SNP array在胎儿超声异常及孕妇不良生育史产前诊断中的应用研究[J].中华医学遗传学杂志,2017,34(2):127-129.
[11]Orhant L,Anselem O,Fradin M,et al.Droplet digital PCRcombined with minisequencing,a new approach to analyze fetal DNA from maternal blood:application to the non‐invasive prenatal diagnosis of achondroplasia[J].Prenatal Diagnosis,2016,36(5):397-406.
[12]刘宁,赵德华,李晓乐,等.PTPS基因检测在6丙酮酰四氢蝶呤合成酶缺乏症出生后诊断和产前诊断中的应用[J].中华妇产科杂志,2016,51(12):890-894.
[13]丁圆,李东晓,刘玉鹏,等.黏多糖贮积症Ⅶ型1例患儿的临床与GUSB基因分析及其同胞的产前诊断[J].中华实用儿科临床杂志,2016,31(8):604-608.
[14]林洋洋,徐婉芳,刘彦慧,等.大规模平行测序技术在产前无创检测胎儿染色体异常中的临床应用[J].中国优生与遗传杂志,2016,1:52-54.
[15]陈梅环,黄海龙,王燕,等.东南亚缺失型遗传性持续性胎儿血红蛋白增多症伴β地中海贫血一个家系的产前诊断[J].中国实验血液学杂志,2017,25(4):1142-1146.