胎儿染色体非整倍体无创基因检测试剂盒的临床验证
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摘要
目的验证胎儿染色体非整倍体(T21、T18、T13)基因检测试剂盒(联合探针锚定连接测序法)在临床检测上的准确性和有效性。方法选择广东省东莞市妇幼保健院的孕妇外周血样本,采用胎儿染色体非整倍体(T21、T18、T13)基因检测试剂盒通过联合探针锚定连接测序法检测,并将结果与临床诊断金标准染色体核型分析结果或出生随访结果作对比。对灵敏度、特异性、总符合率和一致性系数Kappa值进行评价,以验证该试剂盒在临床检测上的准确性和有效性。结果共完成有效样本389例,测序法检出T21阳性6例,T18和T13均未检出阳性,与染色体核型分析结果相符,检测为阴性共383例,经电话随访至出生未发现非整倍体胎儿。统计分析结果显示,测序法与核型分析法总符合率为100%,Kappa=1(≥0.8),其中检测T21的灵敏度、特异性、阳性预测值、阴性预测值均为100%;检测T18和T13的特异性、阴性预测值均为100%。结论采用试剂盒通过联合探针锚定连接测序法检测T21、T18、T13的结果准确度高且方法安全,可以在临床上推广使用。
Objective To verify the accuracy and effectiveness of a genetic detection kit for noninvasive fetal trisomy( T21,T18 and T13,based on combinatorial probe-anchor ligation sequencing,a product of BGI Biotechnology Limited Co.) in clinical practice. Methods Pregnant women who underwent prenatal diagnosis in our hospital were recruited in this study. Maternal peripheral blood samples were collected from each participant,and then processed in accordance with the kit instructions followed by combinatorial probe-anchor ligation sequencing. The results were compared with the results from prenatal karyotyping confirmation and baby born. The sensitivity,specificity,overall compliance rate and consistency coefficient Kappa values were evaluated to verify the accuracy and validity of the kit in the clinical testing.Results A total of 389 valid samples was tested using this genetic testing kit. Six pregnant women were found carrying trisomy 21 fetuses by sequencing,and none of them carrying trisomy 18 or 13 fetus,which consistent with the results of karyotyping. The other 383 women were negative,which was further validated by telephone follow-up after baby born. Statistical analysis showed that the total coincidence rate of our sequencing and karyotyping was 100% and the Kappa = 1( ≥ 0. 8), indicating that the sensitivity,specificity,positive predictive value and negative predictive value of trisomy 21 were 100%, and the specificity,negative predictive value of trisomy 18 and trisomy 13 were 100%. Conclusion This genetic detection kit for noninvasive fetal trisomy is of high accuracy and security,and should be worth of extensive application in clinic.
Objective To verify the accuracy and effectiveness of a genetic detection kit for noninvasive fetal trisomy( T21,T18 and T13,based on combinatorial probe-anchor ligation sequencing,a product of BGI Biotechnology Limited Co.) in clinical practice. Methods Pregnant women who underwent prenatal diagnosis in our hospital were recruited in this study. Maternal peripheral blood samples were collected from each participant,and then processed in accordance with the kit instructions followed by combinatorial probe-anchor ligation sequencing. The results were compared with the results from prenatal karyotyping confirmation and baby born. The sensitivity,specificity,overall compliance rate and consistency coefficient Kappa values were evaluated to verify the accuracy and validity of the kit in the clinical testing.Results A total of 389 valid samples was tested using this genetic testing kit. Six pregnant women were found carrying trisomy 21 fetuses by sequencing,and none of them carrying trisomy 18 or 13 fetus,which consistent with the results of karyotyping. The other 383 women were negative,which was further validated by telephone follow-up after baby born. Statistical analysis showed that the total coincidence rate of our sequencing and karyotyping was 100% and the Kappa = 1( ≥ 0. 8), indicating that the sensitivity,specificity,positive predictive value and negative predictive value of trisomy 21 were 100%, and the specificity,negative predictive value of trisomy 18 and trisomy 13 were 100%. Conclusion This genetic detection kit for noninvasive fetal trisomy is of high accuracy and security,and should be worth of extensive application in clinic.
引文
[1]张月萍,伍俊萍,李笑天,等.孕中期羊水细胞染色体核型分析及其异常核型发生率的比较[J].中华妇产科杂志,2011,46(9):644-648.DOI:10.3760/cma.j.issn.0529-567x.2011.09.002
[2]Nicopoullos J D,Gilling-Smith C,Almeida P A,et al.The role of sperm aneuploidy as a predictor of the success of intracytoplasmic sperm injection?[J].Hum Reprod,2008,23(2):240-250.DOI:10.1093/humrep/dem395
[3]王斌,陈英耀,石琦,等.我国唐氏综合征的疾病经济负担研究[J].中国卫生经济,2006,25(3):24-26.DOI:10.3969/j.issn.1003-0743.2006.03.008
[4]Williams E S,Cornforth M N,Goodwin E H,et al.COFISH,COD-FISH,Re D-FISH,SKY-FISH[J].Methods Mol Biol,2011,735:113-124.DOI:10.1007/978-1-61779-092-8_11
[5]Leung T Y,Voqel I,Lau T K,et al.Identification of submicroscopic chromosomal aberrations in fetuses with increased nuchal translucency and apparently normal karyotype[J].Ultrasound Obstet Gynecol,2011,38(3):314-319.DOI:10.1002/uog.8988
[6]Vermeesch J R,Fiegler H,de-Leeuw N,et al.Guidelines for molecular karyotyping in constitutional genetic diagnosis[J].Eur J Hum Genet,2007,15(11):1105-1114.
[7]中华人民共和国卫生部.产前诊断技术管理办法[J].中国生育健康杂志,2003,14(2):68-77.DOI:10.3969/j.issn.1671-878X.2003.02.001
[8]ACOG Committee on Practice Bulletins.ACOG Practice Bulletin No.77:screening for fetal chromosomal abnormalities[J].Obstet Gynecol,2007,109(1):217-227.
[9]Driscoll D A,Gross S J,Professional Practice Guidelines Committee.Screening for fetal aneuploidy and neural tube defects[J].Genet Med,2009,11(11):818-821.DOI:10.1097/GIM.0b013e3181bb267b
[10]Benn P A,Fang M,Egan J F,et al.Incorporation of inhibin-A in second-trimester screening for Down syndrome[J].Obstet Gynecol,2003,101(3):451-454.
[11]Malone F D,Canick J A,Ball R H,et al.First-trimester or second-trimester screening,or both,for Down’s syndrome[J].N Engl J Med,2005,353(19):2001-2011.
[12]Nicolaides K H.Nuchal translucency and other first-trimester sonographic markers of chromosomal abnormalities[J].Am J Obstet Gynecol,2004,191(1):45-67.
[13]Platt L D,Greene N,Johnson A,et al.Sequential pathways of testing after first-trimester screening for trisomy 21[J].Obstet Gynecol,2004,104(4):661-666.DOI:10.1097/01.AOG.0000153257.08966.f9
[14]Spencer K,Spencer C E,Power M,et al.Screening for chromosomal abnormalities in the first trimester using ultrasound and maternal serum biochemistry in a one-stop clinic:a review of three years prospective experience[J].BJOG,2003,110(3):281-286.
[15]Wald N J,Rodeck C,Hackshaw A K,et al.First and second trimester antenatal screening for Down’s syndrome:the results of the Serum,Urine and Ultrasound Screening Study(SURUSS)[J].J Med Screen,2003,10(2):56-104.
[16]Wald N J,Watt H C,Hackshaw A K.Integrated screening for Down’s syndrome on the basis of tests performed during the first and second trimesters[J].N Engl J Med,1999,341(7):461-467.DOI:10.1056/NEJM199908123410701
[17]Wapner R,Thom E,Simpson J L,et al.First-trimester screening for trisomies 21 and 18[J].N Engl J Med,2003,349(15):1405-1413.DOI:10.1056/NEJMoa025273
[18]Wright D,Bradbury I,Benn P,et al.Contingent screening for Down syndrome is an efficient alternative to non-disclosure sequential screening[J].Prenat Diagn,2004,24(10):762-766.DOI:10.1002/pd.974
[19]Lo Y M,Chiu R W.Prenatal diagnosis:progress through plasma nucleic acids[J].Nat Rev Genet,2007,8(1):71-77.DOI:10.1038/nrg1982
[20]Zhang H,Gao Y,Jiang F,et al.Non-invasive prenatal testing for trisomies 21,18 and 13:clinical experience from146,958 pregnancies[J].Ultrasound Obstet Gynecol,2015,45(5):530-538.
[2]Nicopoullos J D,Gilling-Smith C,Almeida P A,et al.The role of sperm aneuploidy as a predictor of the success of intracytoplasmic sperm injection?[J].Hum Reprod,2008,23(2):240-250.DOI:10.1093/humrep/dem395
[3]王斌,陈英耀,石琦,等.我国唐氏综合征的疾病经济负担研究[J].中国卫生经济,2006,25(3):24-26.DOI:10.3969/j.issn.1003-0743.2006.03.008
[4]Williams E S,Cornforth M N,Goodwin E H,et al.COFISH,COD-FISH,Re D-FISH,SKY-FISH[J].Methods Mol Biol,2011,735:113-124.DOI:10.1007/978-1-61779-092-8_11
[5]Leung T Y,Voqel I,Lau T K,et al.Identification of submicroscopic chromosomal aberrations in fetuses with increased nuchal translucency and apparently normal karyotype[J].Ultrasound Obstet Gynecol,2011,38(3):314-319.DOI:10.1002/uog.8988
[6]Vermeesch J R,Fiegler H,de-Leeuw N,et al.Guidelines for molecular karyotyping in constitutional genetic diagnosis[J].Eur J Hum Genet,2007,15(11):1105-1114.
[7]中华人民共和国卫生部.产前诊断技术管理办法[J].中国生育健康杂志,2003,14(2):68-77.DOI:10.3969/j.issn.1671-878X.2003.02.001
[8]ACOG Committee on Practice Bulletins.ACOG Practice Bulletin No.77:screening for fetal chromosomal abnormalities[J].Obstet Gynecol,2007,109(1):217-227.
[9]Driscoll D A,Gross S J,Professional Practice Guidelines Committee.Screening for fetal aneuploidy and neural tube defects[J].Genet Med,2009,11(11):818-821.DOI:10.1097/GIM.0b013e3181bb267b
[10]Benn P A,Fang M,Egan J F,et al.Incorporation of inhibin-A in second-trimester screening for Down syndrome[J].Obstet Gynecol,2003,101(3):451-454.
[11]Malone F D,Canick J A,Ball R H,et al.First-trimester or second-trimester screening,or both,for Down’s syndrome[J].N Engl J Med,2005,353(19):2001-2011.
[12]Nicolaides K H.Nuchal translucency and other first-trimester sonographic markers of chromosomal abnormalities[J].Am J Obstet Gynecol,2004,191(1):45-67.
[13]Platt L D,Greene N,Johnson A,et al.Sequential pathways of testing after first-trimester screening for trisomy 21[J].Obstet Gynecol,2004,104(4):661-666.DOI:10.1097/01.AOG.0000153257.08966.f9
[14]Spencer K,Spencer C E,Power M,et al.Screening for chromosomal abnormalities in the first trimester using ultrasound and maternal serum biochemistry in a one-stop clinic:a review of three years prospective experience[J].BJOG,2003,110(3):281-286.
[15]Wald N J,Rodeck C,Hackshaw A K,et al.First and second trimester antenatal screening for Down’s syndrome:the results of the Serum,Urine and Ultrasound Screening Study(SURUSS)[J].J Med Screen,2003,10(2):56-104.
[16]Wald N J,Watt H C,Hackshaw A K.Integrated screening for Down’s syndrome on the basis of tests performed during the first and second trimesters[J].N Engl J Med,1999,341(7):461-467.DOI:10.1056/NEJM199908123410701
[17]Wapner R,Thom E,Simpson J L,et al.First-trimester screening for trisomies 21 and 18[J].N Engl J Med,2003,349(15):1405-1413.DOI:10.1056/NEJMoa025273
[18]Wright D,Bradbury I,Benn P,et al.Contingent screening for Down syndrome is an efficient alternative to non-disclosure sequential screening[J].Prenat Diagn,2004,24(10):762-766.DOI:10.1002/pd.974
[19]Lo Y M,Chiu R W.Prenatal diagnosis:progress through plasma nucleic acids[J].Nat Rev Genet,2007,8(1):71-77.DOI:10.1038/nrg1982
[20]Zhang H,Gao Y,Jiang F,et al.Non-invasive prenatal testing for trisomies 21,18 and 13:clinical experience from146,958 pregnancies[J].Ultrasound Obstet Gynecol,2015,45(5):530-538.