Patel Nayana H, Bhadarka Harsha K, Patel Kruti B, Vaniawala Salil N, Acharya Arpan, Mukhopadhyaya Pratap N, Sodagar Nilofar R
Department of IVF, Akanksha Hospital and Research Institute, Near Shrushti English Medium School, Lambhvel-387310, Anand, Gujarat, India.
SN Gene Lab, Floor, A, Above Kansaar Hotel, President Plaza, Ring Rd, Aveerbhav Society 1, Nanpura, Surat, Gujarat, India.
J Hum Reprod Sci. 2016 Jul-Sep;9(3):200-206. doi: 10.4103/0974-1208.192072.
Genetic profiling of embryos (also known as preimplantation genetic diagnosis) before implantation has dramatically enhanced the success quotient of fertilization (IVF) in recent times. The technology helps in avoiding selective pregnancy termination since the baby is likely to be free of the disease under consideration.
Screening of embryos free from c.1537G>A; p.G513S mutation within the COL4A1 gene for which the father was known in before be in heterozygous condition.
Processing of trophectoderm biopsies was done from twelve embryos for c.1537G>A; p.G513S mutation within the COL4A1 gene. DNA extracted from isolated cells were subjected to whole genome amplification using an isothermal amplification and strand displacement technology. Oligonucleotide primers bracketing the mutation were synthesized and used to amplify 162 base pairs (bp) polymerase chain reaction amplicons originating from each embryo which were subsequently sequenced to detect the presence or absence of the single base polymorphism.
Three out of 12 embryos interrogated in this study were found to be normal while 9 were found to harbor the mutation in heterozygous condition. Implantation of one of the normal embryos following by chorionic villus sampling at 11 week of pregnancy indicated that the baby was free from c.1537G>A; p.G513S mutation within the COL4A1 gene.
Single-cell sequencing is a helpful tool for preimplantation embryo profiling. This is the first report from India describing the birth of a normal child through IVF procedure where a potential pathogenic COL4A1 allele was avoided using this technology.
胚胎植入前的基因分析（也称为植入前基因诊断）近年来显著提高了体外受精（IVF）的成功率。这项技术有助于避免选择性终止妊娠，因为婴儿很可能没有所考虑的疾病。
筛选在COL4A1基因中不存在c.1537G>A；p.G513S突变的胚胎，已知父亲在此基因位点为杂合子状态。
对12个胚胎的滋养外胚层活检样本进行处理，检测COL4A1基因中的c.1537G>A；p.G513S突变。从分离的细胞中提取的DNA使用等温扩增和链置换技术进行全基因组扩增。合成了包围该突变的寡核苷酸引物，并用于扩增来自每个胚胎的162个碱基对（bp）的聚合酶链反应扩增子，随后对其进行测序以检测单碱基多态性的存在与否。
本研究检测的12个胚胎中，有3个被发现正常，9个被发现为杂合子状态携带该突变。其中一个正常胚胎植入后，在妊娠11周时进行绒毛取样，结果显示婴儿不存在COL4A1基因中的c.1537G>A；p.G513S突变。
单细胞测序是植入前胚胎分析的一种有用工具。这是印度的第一份报告，描述了通过IVF程序分娩出一个正常婴儿，其中利用该技术避免了一个潜在的致病性COL4A1等位基因。
