The Feasibility Study Of Deep Sequencing For21-trisomy Diagnosis Of Single Cell

Background Chromosome aneuploidy is the most common chromosomal abnormalitieswhich can lead to embryos stop education and spontaneous abortion. It is also one of thereasons that restrict to the success rate of IVF. Preimplantation genetic screening is usdto select embrys with normal chromosome number and structure for transplantation toexpect to improve the success rate. Currently PGS technologies include: FISHtechnique, CGH and aCGH technology and SNP technology, and all technology exist inits own drawbacks. In order to find a more rapid and comprehensive aneuploidydetection method, we try a new approach to complete the21-trisomy diagnosis of singlecell based on the generation sequencing technology. The establishment of thistechnology platform lay the foundation for further study.Objective To establish the preliminary technology platform of deep sequencingtechnology for21-trisomy detecting of single cell.Methods Aimed to assess the stability of this deep sequencing technology method, itwas used to detect the DNA from peripheral blood of normal women. The MDA ofwhole genome amplification was apply on DNA samples of21–trisomy with differentconcentration and its product was deep sequencing with the purpose to assess thestability of this method. We established the preliminary technology platform of deepsequencing technology for21-trisomy detecting of single cell by using this method todetect21-trisomy of single cell.Result1. Deep sequencing was used to detect the14samples and we found thatstandard deviation of each chromosome are at a low level,with the largest standarddeviation is0.078.2.21chromosome copy number of21-trisomy sample is1.5timesthat of21chromosome copy number of the normal sample, and the copy numbers of therest chromosomes have not yet been found the obvious exception compared with thenormal samples.3. The cut off value of0.5,1and1.5represent the haploid, diploid andtriploid respectively and the sequencing results of21-trisomy sample showed thatchromosome21cut off mean value is1.586and the99%CI is [1.537,1.635]. It is feasibility to use the deep sequencing technology to detect21-trisomy of single cell.Conclusion1. Deep sequencing technology is a stable method that is used to detectDNA from a normal female.2. In this study, by sequencing amplification products ofDNA template with different concentrations, we confirm that deep sequencingtechnology can be used to diagnosis21–trisomy.3. Deep sequencing is a feasiblemethod to detect the21-trisomy of single cell.