要应用长读,第三代测序作为进行结构重排(PGT-SR)和单基因疾病(PGT-M)胚胎检测的一般检查策略的一部分.
前瞻性研究。
体外受精单元。
呈现PGT-SR(n=15)和PGT-M(n=2)的对对。
囊胚活检与易位断点或突变(目标)的分子检测。
详细,父母阶段,围绕目标的单核苷酸多态性(SNP)谱,用于选择信息多态性标记,以简化和促进临床植入前遗传测试(PGT)设计,从而区分携带者和非携带者胚胎。
对于呈现PGT-SR的所有15对夫妇,实现了染色体断点的高清晰度以及紧密定相多态性标记。同样,对于为PGT-M出席的两对夫妇来说,也简单地鉴定了突变周围紧密连锁的信息标记。三对易位t(1;17)(q21;p13),t(3;13)(p25;q21.2),t(12;13)(q23;q22)进行PGT-SR,要求优先鉴定非携带者胚胎进行移植。在选择了一组与断点相关的信息性SNP之后,我们成功地进行了PGT-SR测试,导致所有夫妇的非携带者胎儿持续怀孕。同样,使用基于与亲本突变相关的信息性SNP的测试,一对夫妇用PGT-M治疗枫糖浆尿病,导致无病胎儿持续怀孕。
对于考虑临床PGT的夫妇,目标周围的变异单倍性通过快速选择紧密相关的信息标记用于患者特异性测试设计,从而减少了检查过程.
To apply long-read, third-generation sequencing as a part of a general workup strategy for performing structural rearrangement (PGT-SR) and monogenic disease (PGT-M) embryo testing.
Prospective study.
In vitro fertilization unit.
Couples presenting for PGT-SR (n = 15) and PGT-M (n = 2).
Blastocyst biopsy with molecular testing for translocation breakpoints or mutations (targets).
Detailed, parental-phased, single-nucleotide polymorphism (SNP) profiles around targets for selection of informative polymorphic markers to simplify and facilitate clinical preimplantation genetic testing (PGT) designs that enable discrimination between carrier and noncarrier embryos.
High definition of chromosome breakpoints together with closely phased polymorphic markers was achieved for all 15 couples presenting for PGT-SR. Similarly, for the two couples presenting for PGT-M, tightly linked informative markers around the mutations were also simply identified. Three couples with translocations t(1;17)(q21;p13), t(3;13)(p25;q21.2), and t(12;13)(q23;q22) proceeded with PGT-SR, requesting preferential identification of noncarrier embryos for transfer. Following selection of a set of informative SNPs linked to breakpoints, we successfully performed PGT-SR tests, resulting in ongoing pregnancies with a noncarrier fetus for all couples. Similarly, with the use of tests based on informative SNPs linked to the parental mutations, one couple proceeded with PGT-M for maple syrup urine disease, resulting in an ongoing pregnancy with a disease-free fetus.
For couples contemplating clinical PGT, variant haplophasing around the target reduces the workup process by enabling rapid selection of closely linked informative markers for patient-specific test design.