Setting new standards in IVF

Setting new standards in IVF

The clinical challenge Embryo aneuploidy is known to increase with maternal age, causing increase in miscarriage rates and decrease in live birth rates. Due to this age related reduction in live birth rate many couples turn to IVF. Despite major technical advances in IVF procedures over the last 30 years IVF success rates remain low. According to recent data, women under 35 years can on average expect a live birth in 32-33% of cycles started a rate which reduces to under 15% for women over 40 1. Oocyte aneuploidy and maternal age 80 70 60 50 40 30 20 10 24 chromosome aneuploidy screening Recent RCT evidence 2 shows that 45% of embryos are aneuploid, even in good prognosis patients. Selectively implanting euploid embryos is therefore critical to reducing miscarriage rates, improving success rates and increasing single embryo transfer (SET). 24sure sets the standard for screening aneuploidy with biopsies from all stages of embryo development. 0 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 Maternal age Aneuploidy (%) Live birth (%) Miscarriage (%) 1 HFEA. Fertility treatment in 2010. Trends and figures 2 Yang et al; Journal of Molecular CytoGenetics 2012. 5:25

Significant increase in pregnancy rates The first published randomised prospective IVF study of pre-implantation chromosome analysis using 24sure demonstrates that selectively implanting euploid embryos significantly increases pregnancy rates. Yang et al (2012) report a 65% increase in pregnancy at 20 weeks on a good prognosis patient group. A 69.1% pregnancy rate was achieved after 24sure selection of euploid embryos (n=55) versus 41.7% after standard morphological selection of embryos (n=48). Selection was carried out on fresh day 5 embryos and single embryos were transferred. Results of this study provide direct evidence that 24sure analysis can deliver increases in pregnancy rates even in younger, good prognosis patients who are thought more likely to have favourable IVF outcomes. 24sure comprehensive chromosomal screening has been clinically proven to be the effective means to select the chromosomally normal embryo(s) for transfer to ensure a healthy pregnancy for infertile patients. Dr Zhihong Yang, Pacific Reproductive Center Torrance, CA, USA 65% increase in pregnancy rate in 24sure treatment arm 80% 70% 60% 50% 40% 30% 20% 10% 0% Control morphological selection of embryos Treatment 24sure selection of euploid embryos 20 week pregnancy achieved

Rapid results from single cells 24sure is widely used with cells from all stages of embryo development. Polar bodies 24sure the new standard for preimplantation embryo screening Accurate and reliable technologies are essential for preimplantation genetic screening. 24sure screens all 24 chromosomes and can be used to screen for aneuploidy in first and second polar bodies, blastomere biopsies and trophectoderm biopsies within a fresh IVF cycle. 24sure is automated and objective which ensures that a high degree of confidence can be placed in the results so that only euploid embryos are implanted. Screening with 24sure can be completed in 12 hours and therefore does not interfere with tight schedules of a fresh IVF cycle. Blastomeres Labelling Sample preparation and amplification 3 hours Trophectoderm 3 1 2 hours Reliable results in under 12 hours 24sure uses simple protocols familiar to laboratories experienced in classical molecular techniques. Protocols have been optimised for routine application with minimal tube transfers, documented quality control stages and flexible stop points. 24sure requires minimal specialist hardware and is compatible with low cost, 10 µm, laser scanners. Where high throughput operation is required optional hardware and protocol stages may be substituted to enable plate level operation.

Over 200,000 clinical biopsies processed to date. Independent scientific evidence Multiple papers report positive results, the first RCT has been published and many are ongoing. Over 20 peer-reviewed publications and more than 70 conference abstracts have been published, including the first preimplantation validation study undertaken by ESHRE. See the 24sure publication booklet for more details. We ve provided this service to our patients for just over a year and already about 20 babies have been born and another 50 are on the way. It s a significant breakthrough for patients who are struggling to become pregnant. Dr. Leeanda Wilton, Melbourne IVF, Australia Hybridization 3 1 2 hours Scanning and reporting 2 hours 24sure result 12 hours

A complete data analysis and information management system All BlueGnome microarrays are supplied with complementary licenses to our BlueFuse software which provides a complete solution for the analysis, storage and reporting of 24sure results. BlueFuse supports the complete laboratory workflow, from sample receipt to report sign off, and is trusted by over 200 of the world s leading microarray laboratories to deliver high throughput and scalable microarray services. Sample database BlueFuse is built upon a scalable SQL database in which all sample details, experimental information and results are securely stored. Simple filters, powerful queries and a visual representation of each IVF cycle ensure the right information is available whenever it is required. Automated analysis Microarray images, loaded directly from the scanner, are processed fully automatically, saving time and guaranteeing complete reproducibility. Single click short cuts ensure rapid access to microarray images for quality control purposes.

Clear profiles Powerful visualisation facilities generate profiles from thousands of independent measurements from each 24sure microarray to enable the status of each chromosome to be fully understood and the basis of results confirmed. Profiles are generated against both male and female references in order to enable reliable interpretation sex chromosomes. Concise reports Sophisticated algorithms calculate and call the status of each chromosome as either normal or abnormal, backed by a confidence estimate reflecting the degree of assay noise or ambiguity in the underlying biology. In addition to delivering complete reproducibility and objectivity this approach ensures results are fully comparable between the newest 24sure laboratory and the largest studies published in the literature. At the end of the process an automated cycle report is generated.

The complete package Setting up a new microarray protocol and sourcing all equipment can be time and resource intensive. By providing all consumables and the key capital equipment needed to deliver 24sure screening BlueGnome enables you to confidently report accurate results with minimum start up time. BlueGnome provides a complete suite of products to support the 24sure protocol including; SurePlex DNA Amplification System, labelling systems and reference DNA and two microarray formats optimised for different investigations. 24sure 24sure V3 is designed for the investigation of whole chromosome aneuploidy and uses analytical methods to produce a simple report of aneuploidy in all 24 chromosomes, including the sex chromosomes. 24sure+ is designed for the investigation of sub chromosome imbalances in embryos of reciprocal translocation carriers. 24sure+ includes increased coverage of pericentrometic and subtelomeric regions to ensure the identification and reporting of the smallest derived chromosome fragments. 24sure opens up the exciting possibility to perform PGS not just in complicated patients such as those with poor ovarian reserve, but also egg donors and young women as abnormal embryos are common at any age. Jacques Cohen, PhD. Director, Tyho-Galileo Research Laboratories, Livingston, NJ, USA ClearLab hardware 24sure uses simple protocols that have been optimised for routine application, and the ClearScan Microarray Scanner joins the ClearPack Lite and the ClearHyb Hybridisation and Wash System as part of the suite of validated laboratory hardware optimised for use with BlueGnome s recommended protocols. The combination of validated, optimised protocols and hardware enables laboratories to set up 24sure services with ease. All 24sure products are supplied with illustrated protocols and a full range of technical support and training services based on BlueGnome s experience of establishing microarray based services in cytogenetics laboratories worldwide.

Laboratory training programmes Practical training courses are run over 3 days by experienced BlueGnome scientists to provide you with proven bench techniques to generate high-quality results. A full day is spent on software training and getting the most out of your analyses and reporting. Training courses can be organised onsite at your own lab or at a BlueGnome training facility. Learn the hands-on skills to optimise the amplification and labelling cell biopsies and the hybridization and washing techniques to ensure clear results. Become competent using the latest version of BlueFuse Multi software to complete the analysis, interpretation and reporting of array results. Technical support Rolling 24 hours, ISO 13485 compliant, email and phone support from BlueGnome service centres in Cambridge, Washington and Singapore. We know that chromosome abnormality rates in embryos of young patients are as high as 40%, and increase up to 70% in blastocyst of women >40. Indeed work by Gary Harton while at Reprogenetics showed that if euploid blastocysts are available for transfer, they implant at the same rate regardless of maternal age. Santiago Munne, PhD, Director, Reprogenetics, Livingston, NJ, USA

Biographies Professor Alan Handyside Professor Handyside joined BlueGnome in 2010 as Head of PGD. He was the first chairman of the European Society for Human Reproduction and Embryology (ESHRE) Special Interest Group in Reproductive Genetics and co-founder and first chairman of the ESHRE PGD Consortium. Gary Harton Gary joined BlueGnome earlier this year as the Americas IVF Business Development Manager. He has more than 20 years experience in PGD laboratories and numerous published articles, abstracts and book chapters. Selected publications Yang et al (2012) report a 65% increase in pregnancy at 20 weeks, with a 69.1% pregnancy rate after 24sure selection of euploid embryos (n=55) versus 41.7% after standard morphological selection of embryos (n=48). Results of this study provide direct evidence that 24sure analysis can deliver increase in pregnancy rates even in younger patients who are thought more likely to have favourable IVF outcomes. Results of the ESHRE PGS proof of principle study concluded that the ploidy of embryos could be predicted with acceptable accuracy using 24sure analysis of both polar bodies. 195 zygotes were successfully analysed - 28% normal, 72% aneuploid. Human Reprod. 2011 Sept 15th [Epub ahead of print].

Dr. Tony Gordon Tony joined BlueGnome in 2006 as the first CytoChip arrays were launched and initiated the 24sure business area in 2008. He has 17 years experience in molecular cytogenetic diagnostics. 24sure, successfully used in the initial ESHRE PGS taskforce study to assess the use of microarrays in PGS (June 2010), has now been selected for ESHRE s six centre full RCT trial (May 2011). Hodes-Wertz et al., (2012) analysed 2,282 embryos from couples with idiopathic recurrent pregnancy loss (RPL) using 24sure of which 35% were euploid and 60% aneuploid. In 181 transfer cycles the miscarriage rate was only 6.9% compared to the expected rate of 33.5% in an RPL control population and 23.7% in an infertile control group therefore showing a significant decrease in the miscarriage rate of RPL patients. Fiorentino et al. (2011) analysed 200 samples, 93.5% were successfully amplified, diagnosed and confirmed using STR analysis. Only 16% of samples were chromosomally normal which translated to embryos being suitable for transfer in 17/28 cycles. A pregnancy rate of 70.6% per embryo transfer was achieved in this cohort. Human Reprod. 26(7):1925-35.

The broadest range of applications from Europe s leading microarray supplier 24sure is part of a suite of microarray applications covering pre-implantation, constitutional and cancer cytogenetics. All BlueGnome microarray products share the same workflow and are supported by a common software platform for the analysis and storage of results. BlueGnome Headquartered in Cambridge, UK, BlueGnome s multidisciplinary team of mathematicians, software engineers, molecular biologists and cytogeneticists are dedicated to the investigation of aneuploidy and copy number imbalance using the latest array technologies. BlueGnome s CytoChip is used in over 40 countries to investigate leukaemias and childhood developmental delay. Under the direction of Professor Alan Handyside as Head of PGD, 24sure and 24sure+ have been developed to enable reliable aneuploidy screening of 24 chromosomes in 12 hours. BlueGnome is privately held, profitable and for the last three years has been ranked in the top two fastest growing biotech businesses in the UK (Deloitte Fast 50, 2009, 2010 & 2011) and the fastest growing biotech in UK according to Tech Track 100. BlueGnome received a Queen s Award for Enterprise: International Trade in 2012 Further information For further information please contact us or see www.24suretest.com Head office CPC4, Capital Park Fulbourn Cambridge, CB21 5XE United Kingdom tel: +44 (0) 1223 885900 fax: +44 (0) 1223 885949 email: info@24suretest.com web: www.24suretest.com US office BlueGnome Inc, 11781 Lee Jackson Memorial Highway Suite 190 Fairfax, VA 22033 Toll free: 1-800-418-9656 tel: (703) 385-4634 fax: (703) 385-4636 Representative Office 3 Science Park Drive #02-12/25 Suite 22, The Franklin Science Park I, 118223 Singapore tel: +44 (0) 1223 885900 fax: +44 (0) 1223 885949 SurePlex contains technology developed and manufactured by Rubicon Genomics Inc., Ann Arbor, Michigan, USA. United States and international patents pending. For Research Use Only. Not for use in diagnostic procedures. Version 1.1 October 2012