(086) Determination Of Diagnostic Molecular Markers In Non-Obstructive Azoospermia- Male Infertility Via Next Generation Sequencing Methods

Abstract Introduction 1 in 6 couples are infertile and male factor contributes 49%. As an Urologist, azoospermia especially non obstructive azoospermia(NOA) poses a great clinical challenge for prediction of fertility. The basic understanding any disease starts from its physiology which is determined by genes. With the advent entire human genomic sequencing it has become easier than decades ago to identify genes that causally linked specific disease. Objective Hence Studying molecular basis esp. functional genomics NOA, type infertility with failed spermatogenesis at various stages, can help mechanism spermatogenesis, discover potential molecules further research utility diagnostics, prognostics therapeutics also identification target towards development new contraceptives Methods Sequencing analysis was carried out using TruSeq RNA Library Prep Kit v2andusing Illumina HiSeq 2000.Alignments, transcripts chimeric/transplice molecules, their quantification were performed Kallisto software. Differential expression done Bioconductor package limma. Apart standard data procedures identifying expressed differently novel scoring system been applied rank based on 'Strength Association' (StA) NOA. Up-and down-regulated hierarchically arranged StA scores Testicular biopsies collected 18,7 2 donors, respectively, Obstructive Azoospermia (OA), Congenital Bilateral Absence Vas Deferens (CBAVD), Varicocele conditions, following ethical approved Institutional Biosafety Committee (IBSC) Institute Bioinformatics & Applied Biotechnology (IBAB) Ankur-Manipal hospital, Bengaluru. samples stored RNA-later solution (Ambion, cat # AM7024), according manufacturer's guidelines. extracted Ribo Pure kit (AM1924). Results current study derived first list Strength Association Cluster showed overall gene profiles NOA different compared control normal - irrespective sub-types within each set. More importantly, observations supported decision group testis-samples those other conditions where occurs normally, viz., VA, OA CBAVD. grouping not only enhanced number samples, but helped eventually candidate markers more unique Based 100% consistency differential observed across 18 22 via RNA-seq and/or RT-qPCR results, 16mRNAs being suggested as promising candidates reliable diagnosis Similarly,3chimeric have short-listed diagnostic Conclusions transcriptomic testes donors used corresponding alternatively spliced mRNA-isoforms associated disorder. These listed strength association condition, would suggest most cases, particularly Such association, several forms mRNAs, reported time. If these 19 could be assays, individually or combination other, they avoid need open surgeries detection Disclosure No