P-725 Role of Non-Invasive Preimplantation Genetic Testing- Aneuploidy (NIPGT-A) using spent culture media (SCM) and its concordance with Trophoectoderm (TE) biopsy: A Prospective Cohort Study

Abstract Study question What is the ploidy concordance rate between spent culture media (SCM) and trophectoderm biopsy (TE) using Next Generation Sequencing (NGS) correlation with clinical outcome? Summary answer DNA could be isolated from SCM successful NGS library sequencing. Ploidy per-chromosome of TE vs was 68.38% 85.13%. known already Many challenges are associated biopsy, possibility NiPGT-A cell free (cfDNA) very intriguing, as it will simpler, safer cheaper alternative to gold standard biopsy. The bottlenecks like additional equipment trained embryologist, can substantially reduced. disparities in reported results various studies comparing TE, attributed contamination (maternal, paternal or environment), different laboratory workflow methodologies used for embryo culture, whole genome amplification methods, sequencing methods algorithms analysis. design, size, duration Prospective cohort study conducted tertiary care centre August 2021-June 2022. Ethical Approval taken Institute Ethics Committee. Couples female age ≥ 35 years, one more implantation failures, male factor infertility requiring ICSI, opting elective single euploid blastocyst transfer, were included study. Forty four blastocysts obtained 14 patients who underwent IVF/ICSI cycles, gave consent collection PGT-A respectively. Participants/materials, setting, Individualised ovarian stimulation carried out GnRH antagonist protocol. ICSI done fertilization. Embryos cultured sequential medium. On day 3, after thorough washing separately 10μl micro-droplets. collected on day-5, before extraction, preparation Veriseq PGS kit (Illumina) Illumina MiSeq system. Euploid transfer FET cycle Main role chance Out 44 blastocysts, 31 5 13 6. WGA-DNA 100% successful. Average concentration amplified 34.88 ± 9.56 ng/µl, 32.3 6.84 ng/µl Quality control parameters samples per recommended standards. CNV visualization analysis BlueFuse Multi-Software (Illumina). analysed 26 embryos, which 68.38%, chromosome 85.13% sex-chromosome 73.0%. sensitivity, specificity, PPV, NPV diagnostic accuracy 66.6%, 60%, 87.5%, 30% 65.38% 6 had better 72.7% p > 0.05. good (>BB) poor morphology (<BB)embryos, <BB 83.33% 50%, = 0.16. Per higher 90.5% 80.5%,p= 0.001. Full maternal suspected when aneuploid/mosaic, XY euploid, XX (n 4/14), assessed only embryos. Single 66.6%clinical pregnancy rate. One resulted healthy live-birth, two on-going pregnancies, 30weeks 12weeks period gestation. Limitations, reasons caution Before considering routine practice, each embryology lab needs standardization, implement necessary modifications Labs should ensure that viability not affected try minimize chances external reliably predict rates high confidence. Wider implications findings useful technique assess but presently, combination PGT-A, help prioritizing embryos according their potential. Further required find whether may serve an implemented alone practice future. Trial registration number CTRI/2021/04/033121