| ObjectiveExploring the clinical application valueof chromosomal microarray analysis(CMA)in the detection of fetuses with congenital heart disease(CHD)to provide data support for the the possible application of genome-wide CMA technology in prenatal diagnosis fetal CHD.Methods1.We collected 72 samples of cord blood or amniotic fluid with CHD who accepted invasive prenatal diagnosis(amniocentesis or cordocentesis)for karyotype analysis and CMA from Fujian Provincial Maternity and Children's Hospital during February,2016 to May,2017.The samples were composed of 3 parts which include 29 cases of isolated heart abnormalities and 21 cases of multiple heart abnormalities and 21 cases of extracardiac CHD.2.Using Bio Chain kit to extract the DNA from Amniotic fluid cells;DNA extraction from peripheral blood and cord blood was used by QIAamp DNA Blood Mini Kit.The concentration of the DNA was analysed by Nanodrop 2000.3.A series of treatments for DNA,including digestion,connection,amplification,purification,fragmentation,signals labelled,and hybridization with the chip,dyeing and scanning the chip,were implemented according to the standard experimental of the 750 k chip provided by the Affymetrix Cyto Scan company.4.Using the Ch AS software to analys the scan results(.CEL file).Further analyzing the results of CMA with the related database to judge the properties of CNV.5.Combining the analysis of family and CMA testing of the peripheral blood extracted from fetal parents to make clear the property of CNV in variants of all clinical significance(VOUS).6.Using fluorescence in situ hybridization technique(FISH)to verify a part of pathogenic CNV.7.Using SPSS19.0 software for statistical analysis,analysis of chi-square was applied in the interblock analysis.Only if the P < 0.05,the difference was statistically significant.Results1.17 cases of anomaly was analysed by CMA from all the 72 samples(17/72,23.6%),including 14 cases of current,known pathogenic anomaly(14/72,19.4%),which consist 5 cases of aneuploidies(5/14,35.7%),1 cases of 8 repeats(1/14,7.1%),2 cases of 22q11.1 Micro-repeats(2/14,14.3%),1 cases of 22q11.2 deletion(1/14,7.1%),1 cases of 17p11.2 Micro-repeats(1/14,7.1%),1 cases of 8q21.11 deletion(1/14,7.1%),and 3 cases of CNV changes of submicroscopic structure(3/14,21.4%).2.VOUS were detected in 4 cases of fetal samples.One out of four was detected with pathogenic CNV,1 case without parents back,another two cases were derived from the mother with normal phenotype.Based on this,there will be larger possibility to be in benign.3.The CNV rates of the isolated heart malformation,multi heart malformation,and extracardic associated with intracardiac malformation detected by CMA were 6.9%(2/29),27.3%(6/22)and 28.6%(6/21),respectively.While,the CNV rates detected by karyotype analysis were 6.9%(2/29),13.6%(3/22)and 23.8%(5/21).4.The all 72 samples were performed according to karyotype analysis,two cases were failed to incubate.10 cases of 72 samples were detected with abnormal.It consist with 2 cases of trisomy 21,1 case of trisomy 18,2 cases of marker chromosomes(47,XY,+mar),1 case of chromosome balanced translocation [46,XY,?t(7;8)(q22;q22),?t(8;22)(p12;p12)],and 1 case of pericentric inversion [46,XY,inv(11)(p15q21)dn].Conclusion1.CMA is an effective method for the identification of etiology in fetuses with CHD,which increasing the detection rate by 5.6%(4/72)approximately.2.The detection rate of CNV in CHD combined with additional extrarenal malformations cases was higher than the detection rate of the isolated heart abnormalities cases(28.6% vs 6.9%),and the CNV detection rate in cases with multiple heart abnormalities was higher than in isolated heart abnormalities cases(23.8% vs 6.9%).3.The research results provide the basis for appling the CMA in antenatal CHD detection.We recommend there should be detected by CMA as a common molecular diagnostic technology which CHD fetal cases.This measure would be helpful for clinical genetic counseling. |
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