早孕期自然流产绒毛组织的遗传学检测方法比较
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摘要
目的比较G显带核型分析、荧光原位杂交(fluorescence in situ hybridization,FISH)、多重连接依赖探针扩增(multiplex ligation-dependent probe amplification,MLPA)及染色体微阵列分析(chromosomal microarray analysis,CMA)等技术检测早孕期自然流产样本的结果,探讨这几种遗传学检测技术检测早孕期自然流产样本的临床适用性。方法回顾分析354例进行遗传学检测的早孕期自然流产样本,其中G显带核型分析181例、FISH116例、MLPA43例和CMA32例。结果 181例G显带核型分析成功率86.2%(156/181),检出69例异常核型,阳性率44.2%(69/156),其中染色体数目异常63例,平衡性结构异常6例;116例FISH检测均成功,检出53例异常(13、16、18、21、22、X、Y染色体数目异常),阳性率45.7%(53/116);43例MLPA检测均成功,其中4例异常(13、18、21、X、Y染色体数目异常),阳性率9.3%(4/43);CMA检测的标本32例均成功,异常结果20例,阳性率62.5%,其中染色体数目异常17例,亚显微结构异常(5Mb以下的染色体结构异常)3例。结论和其他方法相比CMA技术快速、成功率高,能够明显提高检测范围和样本的异常检出率,在早孕期自然流产样本遗传学分析中具有较强的临床应用价值。
Objective:To evaluate the clinical application of karyotyping,fluorescence in situ hybridization(FISH),multiplex ligation-dependent probe amplification(MLPA)and chromosomal microarray analysis(CMA)in the test of early miscarriage villus. Methods:Three handred and fifty-four cases of the first trimester spontaneous abortion villus were included in this study. Among these,181 cases were tested by routine karyotyping ananysis,116 cases by FISH,43 cases by MLPA and 32 cases by CMA. Results:Twenty-five of the 181(13.8%)cases that were conduct with karyotyping failed. Sixty-nine cases(44.2%,69/156)were abnormal,including 6 cases of balanced chromosomal translocation. Fifty-three cases of the 116 cases(45.7%)underwent FISH analysis were positive for the numerical analysis of chromosome 13,16,18,21,22 or sex chromosome. For those underwent MLPA,only 9.3%(4/43%)were positive. Of the 32 cases detected by CMA,20 cases were abnormal,which were composed of 17 cases of numerical chromosomal anomaly and 3 cases of submicroscopic(less than 5 Mb)chromosome imbalance,giving rise to a positive rate of 62.5%. Conclusion:Compared with karyotyping,FISH and MLPA,CMA can detect early miscarriage villus with wider detection range and higher detection rate. Given this,it is a more effective detecting method.
Objective:To evaluate the clinical application of karyotyping,fluorescence in situ hybridization(FISH),multiplex ligation-dependent probe amplification(MLPA)and chromosomal microarray analysis(CMA)in the test of early miscarriage villus. Methods:Three handred and fifty-four cases of the first trimester spontaneous abortion villus were included in this study. Among these,181 cases were tested by routine karyotyping ananysis,116 cases by FISH,43 cases by MLPA and 32 cases by CMA. Results:Twenty-five of the 181(13.8%)cases that were conduct with karyotyping failed. Sixty-nine cases(44.2%,69/156)were abnormal,including 6 cases of balanced chromosomal translocation. Fifty-three cases of the 116 cases(45.7%)underwent FISH analysis were positive for the numerical analysis of chromosome 13,16,18,21,22 or sex chromosome. For those underwent MLPA,only 9.3%(4/43%)were positive. Of the 32 cases detected by CMA,20 cases were abnormal,which were composed of 17 cases of numerical chromosomal anomaly and 3 cases of submicroscopic(less than 5 Mb)chromosome imbalance,giving rise to a positive rate of 62.5%. Conclusion:Compared with karyotyping,FISH and MLPA,CMA can detect early miscarriage villus with wider detection range and higher detection rate. Given this,it is a more effective detecting method.
引文
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[11]雷彩霞,张月萍,伍俊萍,等.1437例早孕期自然流产胚胎核型分析[J].生殖与避孕,2014,34(4):328-333.
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[13]Schaeffer AJ,Chung J,Heretis K,et al.Comparative genomic hybridization analysis enhances the detection of aneuploidies and submicroscopic imbalances inspontaneous miscarriages[J].Am J Hum Genet,2004,74(6):1168-74.
[14]Zahir F,Friedman JM.The impact of arraygenomic hybridization on mental retardation research:a review of current technologies and their clinical utility[J].Clin Genet,2007,72(4):271-87.
[2]Warburton D,Susser M,Stein Z,et al.Genetic and epidemiologic investigation of spontaneous abortion:relevance to clinical practice[J].Birth Defects Orig Artic Ser,1979,15(5A):127-136.
[3]Wolf GC,Horger ER.Indications for examination of spontaneous abortion specimens:a reassessment[J].Am J Obstet Gynecol,1995,173(5):1364-1368.
[4]Rubio C,Pehlivan T,Rodrigo L,et al.Embryo aneuploidy screening for unexplained recurrent miscarriage:a minireview[J].Am J Reprod Immunol,2005,53(4):159-165.
[5]Diego-Alvarez D,Ramos-Corrales C,Garcia-Hoyos M,et al.Double trisomy in spontaneous miscarriages:cytogenetic and molecular approach[J].Hum Reprod,2006,21(4):958-966.
[6]朱瑞芳,胡娅莉,许争锋.用绒毛细胞培养法制备染色体[J].江苏医药,2008,34:233-234.
[7]朱瑞芳,朱海燕,王皖骏,等.用荧光原位杂交技术分析自然流产组织中的染色体异常[J].中华医学遗传学杂志,2012,29(5):606-607.
[8]Lomax B,Tang S,Separovic E,et al.Comparative genomic hybridization in combination with flow cytometry improves results of cytogenetic analysis of spontaneous abortions[J].Am J Hum Genet,2000,66:1516-1521.
[9]Bell KA,Van Deerlin PG,Haddad BR,Feinberg RF.Cytogenetic diagnosis of“normal 46,XX”karyotypes in spontaneous abortions frequently may be misleading[J].Fertil Steril,1999,71(2):334-41.
[10]Thein A,Charles A,Davies T,et al.The role of comparative genomic hybridization in prenatal diagnosis[J].Br J Obestet Gynaecol,2001,108(6):642-648.
[11]雷彩霞,张月萍,伍俊萍,等.1437例早孕期自然流产胚胎核型分析[J].生殖与避孕,2014,34(4):328-333.
[12]Rajcan-Separovic E,Qiao Y,Tyson C,et al.Genomic changes detected by array CGH in human embryos with developmental defects[J].Mol Hum Reprod,2010,16(2):125-134.
[13]Schaeffer AJ,Chung J,Heretis K,et al.Comparative genomic hybridization analysis enhances the detection of aneuploidies and submicroscopic imbalances inspontaneous miscarriages[J].Am J Hum Genet,2004,74(6):1168-74.
[14]Zahir F,Friedman JM.The impact of arraygenomic hybridization on mental retardation research:a review of current technologies and their clinical utility[J].Clin Genet,2007,72(4):271-87.