%(6/31) [DOI] /j.issn

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1 [ ] ~37 21~27 31 G %(3/26) 4 / %(6/31) [ ] [ ] R [ ] A [ ] (2016) [DOI] /j.issn Application of array-based comparative genomic hybridization in clinical research of multiple malformations YOU Yan-qin 1, WANG Shu-juan 1, WANG Long-xia 2, LU Yan-ping 1*, REN Yuan 1, LI Ya-li 1 1 Department of Obstetrics and Gynecology, 2 Department of Ultrasonic Diagnosis, General Hospital of PLA, Beijing , China * Corresponding author, luyp301@163.com [Abstract] Objective To analyze the molecular genetics changes of multiple malformations using array-based comparative genomic hybridization (acgh), and evaluate the value of this technique in routine chromosomal karyotype analysis. Methods Thirty one cases were studied of fetal multiple malformations collected by ultrasound examination from Feb to May 2015 in General Hospital of Chinese PLA. The age of gravida ranged from 20 to 37, the gestational time ranged from 21 to 27 weeks. Samples of amniotic fluid and umbilical cord blood were collected, and chromosomal karyotype analysis and acgh were carried out. Results All of 31 samples were successfully detected by G-band chromosomal karyotype analysis and acgh. Among karyotype analysis, 4 samples failed of cultivation, and 1 showed no karyokinesis. Among 26 cases of successful cultivation, 3 were with abnormal karyotype leading to a abnormality rate of 11.54% (3/26). In the acgh analysis, the chromosomal polymorphism with no pathogenicity of microdeletion and microduplication were detected in 21 cases; 6 cases showed abnormal and the abnormality rate was 19.35% (6/31). Conclusions Array-based comparative genomic hybridization technique has the advantages of high resolution and wide coverage, may not only make up for the defect of unable to do karyotyping since failed cultivation or lacking cell vigor, but also reveal the microdeletion and microduplication at submicrostructure level, which is favorable for understanding the reason and providing the evidence of fetal malformation. [Key words] comparative genomic hybridization; abnormalities, multiple; karyotyping [1-2] 2 [ ] [ ] ( ) ( ) [ ] luyp301@163.com 5% 2% 8 92%

2 Med J Chin PLA, Vol. 41, No. 9, September 1, [3] 1992 Kallioniemi [4] (comparative genomic hybridization CGH) 1997 (array-based comparative genomic hybridization acgh) [5] 2010 ( ) acgh 7%~11% [6] acgh 92% 8% FISH [7] acgh ~37 21~ B 22 10ml 5ml 20~30ml 100mg acgh 3ml EDTA g 10min g 10min 80 TIANamp Micro Kit ( ) DNA ng 1 l 6 Loading Buffer 1%DNA Agilent 60K CGH ( ) G %(26/31) %(3/26) 2.2 acgh 31 acgh 4 / %(6/31) 2 XO Mb 22 1 acgh 1 6 acgh G 5~10Mb G 14~21d (fluorescence quantization polymerase chain reaction QFPCR) (multiple ligation-dependent probe amplification MLPA) X,Y acgh

3 acgh Tab.1 The chromosomal karyotyping and acgh result of 31 multiple malformation fetus No. Ultrasound test Chromosomal karyotype acgh results 1 Hydrocephalus, banana cerebellum, meningomyelocele 46,XY No pathogenic chromosomal deficiency/repeat 2 Short limbs, the lower extremities bending 46,XY 3 Fetal ventriculomegaly, left diaphragmatocele, right atresia of external acoustic meatus (Fig.1) 47, xx, t(11;22 (q23;q11), +der(22) t(22;11) (q11;23) A duplication of 18.21Mb in chr11q23.3-q25, including 38 OMIM genes, associated with "11q23.3q25 Syndrome" previously reported in database, involving a series of manifestations like mental retardation, malformation. A duplication of 3.16Mb in chr22q11.1-q11.21, related to "22q11 duplication syndrome" previously reported in DECIPHERS DATABASE, the clinical symptoms includes mental retardation, facial abnormalities. A 18.21Mb duplication in chr11q23.3-q25( ); A 3.16Mb duplication in chr22q11.1-q11.21( ) 4 Short limbs extremely, achondroplasia 46,XY 5 Short limbs, polycystic kidney 46,XX 6 Spinal column, abdominal, limbs 46,XY 7 Hydrocephalus, rachischisis 46,XY No Pathogenic chromosomal deficiency/repeat 8 Bilateral strephenopodia, right arthroclisis 46,XY 9 Deaf, absence of right auricula 46,XX 10 Short limbs extremely, hydrocephalus 46,XY 11 Atrial septal defect, short and bowed limbs, hexadactylia Culture failure 12 Hyposarca, pleural effusion, rachischisis 46,XX 13 Cleft lip and palate, right ureterectasia 46,XY Rachischisis, left hand hexadactylia, double genitalia Multiple malformation(right cleft lip and palate) (Fig.2) Hyposarca, pleural effusion and hydrocephalus (Fig.3) 46,XY Culture failure No split phase 17 Fetal edema, atrial septal defect 46,XX Adeletionof0.92Mbinchr3p26.3,whileaentiremissingofoneoftheChromosomeX, indicating the "XO"; A 0.92Mb deletion in chr3p26.3( ); A 93.26Mb deletion in chrxq11.1-q28( ) A deletion of 0.18Mb in chr7q21.11-q21.12, and a duplication of 0.84Mb in chr11q22.3, while a entire duplication of one of the Chromosome 13, indicating the "13-trisomy"; A 0.18Mb deletion in chr7q21.11-q21.12( ); A 0.84Mb duplication in chr11q22.3( ); A 9.64Mb duplication in chr13q31.1-q31.3( ) 18 Single umbilical artery, right hemivertebrae, right strephenopodia, double polydactyly 46,XY 19 Right polydactylism, left ectodactyle, the absenceofcarpus 46,XX 20 Cleft lip and palate, thickening of nuchal translucency 46,XX No pathogenic chromosomal deficiency/repeat 21 Cleft lip, lateral ventriculomegaly 46,XY No pathogenic chromosomal deficiency/repeat 22 Polydactylism, cleft lip and palate, persistant truncus arteriosus (Fig.4) 47,XN, Hydrocephalus, rachischisis 46,XY 24 Hyposarca, polydactylism 46,XX 25 Heart malformations, large-volume paracentesis 46,XY 26 Cleft lip and palate, left radius absence Culture failure 27 Hydrocephalus, agenesis of corpus callosum 46,XY 28 Cervical lymphatic cyst and hyposarca, pleural effusion, intrauterine death (Fig.5) Culture failure 29 Acromphalus (Fig.6) 47,XN, Multiple malformations (holeprosencephaly, single nostril, umbilical hernia) 46,XX 31 Right lateral ventriculomegaly, fetal growth restriction 46,XY A deletion of 0.18Mb in chr7q21.11-q21.12, a duplication of 0.84Mb in chr11q22.3, and a 2.03Mb duplication in chr15q11.1-q11.2, while a entire duplication of one of the chromosome 13, indicating the "13-trisomy"; A 0.18Mb deletion in chr7q21.11-q21.12( ); A 0.84Mb duplication in chr11q22.3( ); A 2.03Mb duplication in ch r15q11.1-q11.2( ); A 95.76Mb duplication in chr13q11 -q34( ) A deletion of 0.18Mb in chr7q21.11-q21.12, and a 2.03Mb duplication in chr15q11.1-q11.2, while a entire missing of one of the Chromosome X, indicating the "XO"; A 0.18Mb deletion in chr7q21.11-q21.12( ); A 2.03Mb duplication in chr15q11.1-q11.2( ); A 93.26Mb deletion in chrxq 11.1-q28( ) A duplication of 0.50Mb in chr2p16.1( ), a duplication of 0.73Mb in chr11q22.3( ), while a entire duplication of one of the chromosome 18, indicating the "18-trisomy"; A 0.5Mb duplication in chr2p16.1( ); A 0.84Mb duplication in ch r11q22.3( ); A 59.36Mb duplication in chr18q11.1 -q23( )

4 Med J Chin PLA, Vol. 41, No. 9, September 1, acgh Fig. 1 Karyotyping and acgh results of case 3 A. Karyotyping result: 47,xx,t(11;22(q23;q11),+der(22)t(22;11)(q11;23); B. acgh result: a 18.21Mb duplication in chr11q23.3 -q25( ), a 3.16Mb duplication in chr22q11.1-q11.21( ) <1kb (copy number variation CNV) [8] Fiorentino [9] acgh acgh 2012 Wapner [10] 4406 acgh acgh

5 acgh Fig.2 acgh results of case 15 A deletion of 0.92Mb in chr3p26.3, while an entire missing of one of the Chromosome X, indicating the "XO" 3 16 acgh Fig.3 acgh results of case 16 A deletion of 0.18Mb in chr7q21.11-q21.12, and a duplication of 0.84Mb in chr11q22.3, while an entire duplication of one of the Chromosome 13, indicating the "13-trisomy" 6.0% 1.7% acgh acgh 1%~2% 0.2% [11] 3.2 acgh 31

6 Med J Chin PLA, Vol. 41, No. 9, September 1, 2016 Ȟ755 ప4Ȟ ҷ22 ᒎ㶔 acgh Fig.4ȞDeformity and acgh results of case 22 ȞȞA. Deformity expression: polydactylism, cleft lip and palate, persistent truncus arteriosus; Karyotyping result: 47,XN,+13; B. acgh result: a deletion of 0.18Mb in chr7q21.11-q21.12, a duplication of 0.84Mb in chr11q22.3, and a 2.03Mb duplication in chr15q11.1-q11.2, while an entire duplication of one of the chromosome 13, indicating the "13-trisomy" ప5Ȟ ҷ28 ᒎ㶔 acgh Fig.5ȞDeformity and acgh results of case 28 ȞȞA. Deformity expression: Cervical lymphatic cyst and hyposarca, pleural effusion, intrauterine death. Karyotyping result: culture failure; B. acgh result: a deletion of 0.18Mb in chr7q21.11-q21.12, and a 2.03Mb duplication in chr15q11.1-q11.2, while an entire missing of one of the chromosome X, indicating the "XO"

7 acgh Fig.6 Deformity, karyotyping and acgh results of case 29 A. Deformity expression: acromphalus. Kayrotyping result: 47,XN, +18; B. acgh result: a duplication of 0.50Mb in chr2p16.1( ), a duplication of 0.73Mb in chr11q22.3( ), while an entire duplication of one of the chromosome 18, indicating the "18-trisomy" G acgh acgh 31 G ,xx,t(11;22) (q23;q11) +der(22)t(22;11)(q11;23) 47,XN,+13 47,XN, %(3/26) 3 acgh 22 acgh Mb Mb Mb acgh Mb Mb acgh Mb Mb (nuchal translucency NF) 11 11q23.3q25 Syndrome 38 OMIM Mb DECIPHERS 22q11 Turner 1/2500 X 45X( 50% ) ( 45,X/46,XX 45X/47,XXX) acgh 2 45X acgh

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