RESUMEN
Background: This study aimed to perform preimplantation genetic testing (PGT) for a female Coffin-Lowry Syndrome (CLS) patient with a de novo mutation (DNM) in RPS6KA3. It was challenging to establish the haplotype in this family because of the lack of information from affected family members. Hence, we explored a new and reliable strategy for the detection of the DNM in PGT, using Oxford Nanopore Technologies (ONT) and the MARSALA platform. Methods: We performed whole-exome sequencing (WES) on the proband and confirmed the pathogenic mutation by Sanger sequencing. The proband then underwent PGT to prevent the transmission of the pathogenic mutation to her offspring. We diverged from the conventional methods and used long-read sequencing (LRS) on the ONT platform to directly detect the mutation and nearby SNPs, for construction of the haplotype in the preclinical phase of PGT. In the clinical phase of embryo diagnosis, the MARSALA method was used to detect both the SNP-based haplotype and chromosome copy number variations (CNVs), in each blastocyst. Finally, a normal embryo was selected by comparison to the haplotype of the proband and transferred into the uterus. Sanger sequencing and karyotyping were performed by amniocentesis, at 17 weeks of gestation, to confirm the accuracy of PGT. Results: Using WES, we found the novel, heterozygous, pathogenic c.1496delG (p.Gly499Valfs*25) mutation of RPS6KA3 in the proband. The SNP-based haplotype that was linked to the pathogenic mutation site was successfully established in the proband, without the need for other family members to be tested with ONT. Eight blastocysts were biopsied to perform PGT and were assessed with a haplotype linkage analysis (30 SNP sites selected), to give results that were consistent with direct mutation detection using Sanger sequencing. The results of PGT showed that three of the eight blastocysts were normal, without the DNM. Moreover, the patient had a successful pregnancy, after transfer of a normal blastocyst into the uterus, and delivered a healthy baby. Conclusion: The ONT platform, combined with the MARSALA method, can be used to perform PGT for DNM patients without the need for other samples as a reference.
RESUMEN
OBJECTIVE: To assess usefulness of serum inhibin B (INHB) measurement for evaluation of ovarian function. METHODS: Serum INHB level on day 3 of menstrual cycle was determined by enzyme labeled immunosorbent assay (ELISA) in 96 cases of in vitro fertilization and embryo transfer (IVF-ET). The patients were classified into 3 groups including (1) poor response (n = 6), normal response (n = 72), and over response (n = 18) according to their response to ovary stimulation. In addition, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E(2)) levels were also determined by chemiluminescent microparticle immunoassay in these patients. Evaluation of the INHB measurement and related factors was performed by statistical analysis. RESULTS: Serum INHB levels in poor, normal and over-response groups were (28 +/- 20), (85 +/- 42), (92 +/- 34) pg/ml; FSH levels were (11.9 +/- 5.3), (7.5 +/- 2.6), (7.2 +/- 1.7) U/L; E(2) levels on day 3 of human chorionic gonadotropin (hCG) administration were (2558 +/- 2108), (9366 +/- 4472), (18 392 +/- 9655) pmol/L; numbers of retrieved oocytes per cycle were (0.6 +/- 0.4), (8.7 +/- 3.6), (14.3 +/- 2.9); top grade embryos were (0.4 +/- 0.3), (3.8 +/- 1.9), (4.6 +/- 1.7); pregnancy rates were 16.7%, 36.1%, 61.1%, respectively. INHB level was negatively correlated to FSH (r = -0.222, P < 0.05) and FSH/LH (r = -0.371, P < 0.05); while positively correlated to E(2) on the day of hCG administration (r = 0.336, P < 0.05), number of retrieved oocytes (r = 0.404, P < 0.05), number of quality embryos (r = 0.323, P < 0.05) and pregnancy rate (r = 0.246, P < 0.05), respectively. CONCLUSIONS: INHB test may reflect the ovarian reserve which is of clinic importance in the guidance of controlled ovarian hyperstimulation.
