BlueFuse Multi Software Research Articles

Impact of the number of previous miscarriages on the clinical outcome of recurrent miscarriage couples undergoing comprehensive chromosome screening (CCS)

To assess the impact of the number of previous miscarriages in couples with ≥2 miscarriages of unknown etiology. Retrospective observational study. A total of 132 CCS cycles were included in women ≤38 years of age, from January 2010 to December 2012. Day-3 embryo biopsies were performed and embryo transfer was scheduled on day-5. A single blastomere from each embryo underwent Whole Genome Amplification (WGA) with Sureplex DNA Amplification System (BlueGnome Ltd, Cambridge, UK). WGA products and genomic DNA used as control were labeled with Cy3 and Cy5 fluorophores. Both labeled DNAs were co-hybridized overnight in the 24sure platform (BlueGnome Ltd, Cambridge, UK). Slides were scanned and analyzed by BlueFuse Multi software (BlueGnome Ltd, Cambridge, UK). Pearson's correlation (p<0.05) was performed using SPSS Statistics for Windows, Version 17.0 (Chicago: SPSS Inc.). Results according to the number of previous miscarriages are shown in the table. No correlation was observed for the described variables.Tabled 12 miscarriages3 miscarriages4 miscarriages≥ 5 miscarriagesNo. of cycles5053236% Transfers88.077.369.6100% Abnormal embryos56.666.465.250.0% Complex aneuploidies*Embryos with aneuploidies for more than one chromosome.11.116.321.520.8% Partial aneuploidies7.34.85.90% Chaotic pattern13.918.310.420.8Pregnancy rate/transfer54.558.568.766.7Miscarriage rate4.220.8012.5Implantation rate50.742.466.740.0Pregnancy rate/cycle48.045.355.066.7* Embryos with aneuploidies for more than one chromosome. Open table in a new tab High pregnancy and implantation rates were achieved with CCS in couples experiencing repeated miscarriages of unknown etiology. No correlation was found with the number of previous miscarriages and the percentage and type of chromosome abnormalities. Reproductive outcome was not affected by the number of previous miscarriages.

A preliminary study on the application of array comparative genomic hybridization for preimplantaion genetic diagnosis

To assess the value of array comparative genomic hybridization (array CGH) technique for preimplantation genetic diagnosis (PGD). Array CGH was performed on three types of cells, which included 3-5 cells isolated from B2/C38/A1 embryonic stem cell lines, single cells isolated from two discarded normal fertilized embryos, and 10 blastocysts biopsied from 5 couples undergoing PGD for chromosomal translocations. For the 10 blastocysts, 8 were abnormal embryos, 1 appeared to be normal but showed arrested development, and 1 embryo was without any fluorescence signals. 24sure V3 or 24sure+ array chips were applied for CGH analysis. The results were analyzed with a BlueFuse Multi software. (1) The results of cells from B2/C3/A1 embryo stem cells by array CGH were consistent with karyotyping analysis. (2) For the 6 single cell samples from two discarded embryos, 2 blastomeres from one embryo were diagnosed as with aneuploidy and a normal karyotype, respectively. Two out of 4 blastomeres biopsied from another embryo were normal, whilst the remaining two were diagnosed with aneuploidies of -22 and +13. Repeated detection with 24sure+ array was consistent with the 24sure V3 result. (3) Ten cell masses from 10 embryos in PGD cycles were successfully analyzed with array CGH, among which four were confirmed with fluorescence in situ hybridization (FISH) on day 3. In two of them, array CGH confirmed FISH diagnosis. For the remaining two, additional aneuploidies for chromosomes not tested by FISH were discovered by array CGH. Another embryo diagnosed as no signal by FISH was found to have trisomy 13 by array CGH. The remaining 5 embryos also showed discordant results by FISH and array CGH. One embryo from a Robertsonian translocation carrier was found to have monosomy 13 by FISH but trisomy 14 and additional aneuploidies by both 24sure V3 and 24sure+ chips. One embryo with many fragments and arrested development by D5 showed discordant results by FISH and array CGH. However, the FISH and array CGH results for other two embryos from this reciprocal translocation carrier were consistent. Three embryos with inconsistent results by FISH and array CGH had a chromosomal translocation involving q11 region. Array CGH is useful for PGD applications for its capability to detect structural chromosomal abnormalities through screening of aneuploidies. However, the 24sure V3 array may not suit detection of translocations with breakpoints close to the q11 region of chromosomes.

Abstract 314: Distinctive genomic signature patterns of common Hematological malignancies uncovered by chromosomal microarray analysis (CMA) using a cancer-specific microarray

Abstract The application of chromosomal microarray analysis (CMA) in cancer research has produced a wealth of useful information about copy number alterations (CNAs) and their implications in cancer classification, disease progression, therapy response, and patient outcome. However, only a handful of clinical laboratories are offering CMA for cancer diagnosis. This is largely due to the lack of effective diagnostic standards and guidelines for cancer applications, and the absence of a cancer microarray database to aid in post-analytic interpretation. We have designed a combined targeted-/whole-genome array specific for cancer using the microarray CGH format. Approximately 20,000 high quality oligonucleotide probes were selected from the Agilent Tech. (Santa Clara, CA) eArray system to target all exons and exon/intron boundaries of more than 500 cancer genes, over 100 known cancer-associated genomic regions and all subtelomeric chromosome regions. Intervals between aforementioned genes or regions were filled relatively evenly with oligonucleotide probes to cover the whole genome. Using this custom designed array, we studied 250 cases of hematological malignancies and 30 normal bone marrow or blood controls. In addition to confirming and clarifying cytogenetic and FISH results, the array revealed many previously unknown CNAs, including intra-gene deletions and duplications in patients with normal or abnormal karyotype. Many apparently balanced translocations were found to harbor cryptic CNAs at or near the breakpoints. Many CNAs displayed strong association with specific category of malignancies. GeneSpring Hierarchical Clustering analysis showed distinctive genomic signature patterns for different types or sub-types of haematological cancers. Using BlueFuse Multi software, we were able to create a few disease classification/decision tracks, which are of great clinical significance in post-analytic interpretation and genotype-phenotype correlation for cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 314. doi:10.1158/1538-7445.AM2011-314