OBJECTIVE: The cytogenetic analysis for earlier detection of fetal chromosome aneuploidies is performed from chorionic villus using either long-term culture or direct chromosome preparation. To analyze the cause of pregnancy loss, we also attempt the cytogenetic study in product of conception(POC) using chorionic villi or fetal tissue. But the failure of analysis often occurs in direct preparation of villus cells and product of conception(POC). We studied to evaluate the clinical usefulness of FISH in uncultured chorionic villus cells of culture-failed cases. METHODS: According to the patient's indication, we performed FISH for chromosome 18, 21, X and Y in chorionic villi as well as POC and compared FISH results with their chromosomal studies. RESULTS: We found one trisomy 18 and one trisomy 21 in Chorionic Villus Sampling and one trisomy 18 and one monosomy X(45, X) in POC. The averages for accuracy of FISH were 83-91% and all cases are represented consistent results with their chromosomal studies. Among them, we could analyze using FISH only in 5 cases of culture failure including one case of monosomy X in POC. CONCLUSION: We could detect aneuploidy with uncultured chorionic villus cells in case of culture failure, using FISH, it may be the potential method to assist the cytogenetic study.
Aneuploidy ; Chorion* ; Chorionic Villi Sampling ; Chorionic Villi* ; Chromosomes, Human, Pair 18 ; Cytogenetic Analysis ; Cytogenetics ; Down Syndrome ; Female ; Fetus ; Monosomy ; Pregnancy ; Trisomy ; Turner Syndrome

OBJECTIVETo detect the relationship between chromosomal anomalies and the pathogenesis, development and prognosis of ovarian carcinoma.

METHODSThirty-six specimens of ovarian carcinoma (n=12), ovarian benign tumor (n=12), and normal ovary (n=12) were examined by fluorescence in situ hybridization (FISH).

RESULTSTwelve cases of mutations, including trisomy 8, monosomy 8 or tetraploid 8 chromosomal anomalies, were found in the group of ovarian carcinoma, making up 100% (12/12). Three cases of trisomy 8 chromosomal anomalies were found in the group of ovarian benign tumor, accounting for 25% (3/12). No anomaly was found in the normal group. There were significant differences between the three groups, P<0.001.

CONCLUSIONThe above anomalies of chromosome 8 are significantly associated with the pathogenesis and development of ovarian carcinoma. The anomalies may occur in the early stage of the carcinoma, and may be significantly associated with the pathological differentiation and clinical stage of the case.

Adolescent ; Adult ; Aged ; Aneuploidy ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Middle Aged ; Monosomy ; Ovarian Neoplasms ; genetics ; Trisomy

Comparative genomic hybridization (CGH) can now be applied to detect the origin of extra or missing chromosomal material in cases with common unbalanced aberrations and in prenatal investigations. This method has been used in 13 cases of fetal samples for this study; 3 for amniocytes, 2 for cord blood and 8 for abortus tissues. These samples were previously subjected to GTG-banding. Our study showed aneuploidy in 8 cases, and partial monosomy, partial trisomy or marker chromosome in the remaining 5. The CGH disclosed further small genetic imbalances in 4 of all 13 cases: a prenatal sample showing del(20)(q13) by GTG confirmed a loss of the segment 20p13-pter by CGH; a marker chromosome manifested normal CGH profile; chromosome der(?)(?;15) found in an abortus sample by GTG turned out to be a loss of 15pter-q14 (partial monosomy) and a gain of 10pter-q22 (partial trisomy); the der(15) shown by GTG represented partial trisomy of 3q24-qter. These findings show that CGH is very useful and efficient for cytogenetic investigations of clinical cases.
Aneuploidy ; Chromosome Aberrations* ; Chromosome Deletion ; Comparative Genomic Hybridization* ; Cytogenetics ; Fetal Blood ; Trisomy

OBJECTIVETo explore the limitation of non-invasive prenatal testing (NIPT) technique through analyzing two false negative cases.

METHODSChromosomal karyotyping analysis was performed on umbilical cord blood sample derived from case 1 at 24 weeks' gestation and peripheral blood sample derived from the neonate of case 2. Placental tissues of case 1 and peripheral blood sample of case 2 were also analyzed by high-throughput sequencing for copy number variations (CNVs).

RESULTSFor case 1, analysis of fetal umbilical cord blood sample showed a translocation type of trisomy 21, i.e., 46,XY,der(21;21)(q10;q10),+21. There were no obvious abnormalities detected at or near the center of the fetal surface and matrix surface of the placenta. High-thoroughput sequencing showed Chr13:(33 840 001 - 115 100 000)×3[60%]/46,XY[40%] at the edge of the placenta, Chr13:(34 080 001-115100000)×3[54%]/46,XY[46%] at the edge of placenta matrix surface, and trisomy 21 in the umbilical cord tissue. For case 2, analysis of the neonatal peripheral blood sample showed a karyotype of 46,XY,del(18)(q22), which revealed a microdeletion in chromosome 18. High-throughput sequencing of the maternal peripheral blood sample stored during pregnancy confirmed it to be chr18: (62 910 000 - 78 020 000)×1 with 15.1 Mb deletion in the fetus. The neonate was therefore diagnosed with partial monosomy of chromosome 18.

CONCLUSIONFalse negative results of NIPT are related with the fraction of circulating cell-free fetal DNA in the maternal serum. NIPT has limitations in detecting fetal chromosomal microdeletion and confined placenta mosaicisms. Routine ultrasound scan is necessary for pregnant women with low-risk indicated by NIPT.

Adult ; Aneuploidy ; Chromosomes, Human, Pair 18 ; Diagnostic Errors ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Karyotyping ; Monosomy ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVE: To explore whether it is necessary to choose NIPT-plus for the prenatal screening of pregnant women. METHODS: The results of NIPT and NIPT-plus sequencing data, fetal DNA concentration, prenatal diagnosis and pregnancy outcome of 50 pregnant women were compared. RESULTS: Compared with NIPT, NIPT-plus attained similar fetal DNA concentration and a 4.4-fold increase in sequencing data. NIPT was able to detect 4 cases of 21-trisomy, 2 cases of 18-trisomy, and 9 cases of sex chromosome aneuploidies (SCAs) signaled by NIPT-plus, but missed one 18-trisomy, and failed to detect rare chromosome aneuploidies (RCAs) and microdeletion/microduplication syndromes (MMS). The PPVs of NIPT-plus for 21-trisomy, 18-trisomy, SCAs, MMS and RCAs were 100%, 100%, 44.4%, 30.4% and 0%, respectively. And those of NIPT for 21-trisomy, 18-trisomy, and SCAs were 100%, 100%, and 44.4%, respectively. CONCLUSION It is necessary for pregnant women to select NIPT-plus to improve the detection rate of common trisomies, SCAs and disease-specific MMS, therefore reduce the occurrene of birth defect.
Aneuploidy ; Female ; Humans ; Pregnancy ; Pregnant Women ; Prenatal Diagnosis ; Trisomy ; Trisomy 13 Syndrome ; Trisomy 18 Syndrome

Constitutional trisomy 8 is a relatively rare aneuploidy; most identified cases are mosaic with a normal cell line. The phenotype is highly variable from apparently normal to severe disability. The proportion of abnormal cells is dramatically different between tissues and the severity of the phenotype is not directly related to the level of mosaicism. Therefore, it is very difficult to provide a definitive prognosis. We report here a case of constitutional trisomy 8 mosaicism with agenesis of the corpus callosum, congenital heart disease and micrognathia. The trisomy 8 cell line was not detected by prenatal cytogenetic study. This is the fourth reported case of constitutional trisomy 8 mosaicism in Korea.
Aneuploidy ; Cell Line ; Corpus Callosum ; Cytogenetics ; Heart Defects, Congenital ; Korea ; Mosaicism* ; Phenotype ; Prognosis ; Trisomy*

Constitutional trisomy 8 mosaicism (CT8M) is a relatively rare aneuploidy in humans with characteristic phenotypes including typical craniofacial feature (such as deformed skull, prominent forehead, low-set and/or dysplastic ears), skeletal malformation, cardiac anomaly, renal malformation, cryptochidism, varying degree of developemental delay. Due to the extremely variable phenotypic and cytogenetic expression, CT8M has gone undiagnosed in certain patients. We report a 28-year-old women with secondary amenorreha without characteristic CT8M phenotype. Chromosomal analysis showed a CT8M (47,XX,+8[9]/46,XX[41]).
Adult ; Aneuploidy ; Chromosomes, Human, Pair 8 ; Cytogenetics ; Female ; Forehead ; Humans ; Mosaicism ; Phenotype ; Skull ; Trisomy ; Uniparental Disomy

PURPOSE: We developed and validated a fetal trisomy detection method for use as a noninvasive prenatal test (NIPT) including a Clinical Laboratory Improvement Amendments (CLIA)-certified bioinformatics pipeline on a cloud-based computing system using both Illumina and Life Technology sequencing platforms for 221 Korean clinical samples. We determined the necessary proportions of the fetal fraction in the cell-free DNA (cfDNA) sample for NIPT of trisomies 13, 18, and 21 through a limit of quantification (LOQ) test. MATERIALS AND METHODS: Next-generation sequencing libraries from 221 clinical samples and three positive controls were generated using Illumina and Life Technology chemistries. Sequencing results were uploaded to a cloud and mapped on the human reference genome (GRCh37/hg19) using bioinformatics tools. Based on Z-scores calculated by normalization of the mapped read counts, final aneuploidy reports were automatically generated for fetal aneuploidy determination. RESULTS: We identified in total 29 aneuploid samples, and additional analytical methods performed to confirm the results showed that one of these was a false-positive. The LOQ test showed that the proportion of fetal fraction in the cfDNA sample would affect the interpretation of the aneuploidy results. CONCLUSION: Noninvasive chromosome examination (NICE), a CLIA-certified NIPT with a cloud-based bioinformatics platform, showed unambiguous success in fetus aneuploidy detection.
Aneuploidy* ; Computational Biology ; DNA ; Fetus* ; Genome ; High-Throughput Nucleotide Sequencing ; Humans ; Prenatal Diagnosis ; Trisomy

BACKGROUND: Serological prenatal screening tests are widely used to detect fetal chromosomal abnormalities such as Down and Edward syndromes. After determining the presence of fetal cell-free DNA in maternal blood, the non-invasive prenatal test (NIPT) coupled with next-generation sequencing has been performed in other countries, therefore, we developed a domestic NIPT technology. METHODS: The results of genomics-based NIPT performed between April and May, 2015 were analyzed. Maternal blood samples were collected in a specific Cell-Free DNA BCT tube. The samples were then massively sequenced using MiSeq and NextSeq 500 (Illumina Inc., USA) using LabGenomics laboratory-developed libraries. Chromosomal abnormalities were analyzed using a bioinfomatics algorithm. RESULTS: A total of 464 cases were analyzed. The samples of 12 subjects had to be collected again because of a low fetal DNA fraction in the initially obtained samples. Among the 456 cases for which fetal genome results were obtained, 436 had a low risk of trisomy, 12 had a high risk for Down syndrome, two had a high risk for Edward syndrome, and four had sex chromosomal aneuploidy, showing that the positive percentage of chromosomal abnormalities was 4.4%. All 12 cases with high risk for Down syndrome were confirmed as having trisomy 21 by amniocentesis. CONCLUSIONS: Our laboratory-developed genomics-based NIPT showed high positive predictive value, therefore, NIPT may be replaced by our own developed method.
Amniocentesis ; Aneuploidy ; Chromosome Aberrations ; DNA ; Down Syndrome ; Genome ; Prenatal Diagnosis ; Trisomy

Complete trisomy 9 is a rare chromosomal aneuploidy in live born infants. The majority of cases of trisomy 9 end in spontaneous abortion in the first trimester. Clinical finding of co-mplete trisomy 9 demonstrate multiple organ abnormalities in the craniofacial, cardiovascular, skeletal, genitouronary systems. We report a fetus with Dandy Walker syndrome which was diagnosed prenatally and was subsequently found to have a complete trisomy 9.
Abortion, Spontaneous ; Aneuploidy ; Dandy-Walker Syndrome* ; Female ; Fetus ; Humans ; Infant ; Pregnancy ; Pregnancy Trimester, First ; Trisomy*