Hearing impairment (HI) caused by mutations in the connexin-26 gene (GJB2) accounts for the majority of cases with inherited, nonsyndromic sensorineural hearing loss. Due to the illegality of the abortion of deaf fetuses in Islamic countries, preimplantation genetic diagnosis (PGD) is a possible solution for afflicted families to have a healthy offspring. This study describes the first use of PGD for GJB2 associated non-syndromic deafness in Iran. GJB2 donor splicing site IVS1+1G>A mutation analysis was performed using Sanger sequencing for a total of 71 Iranian families with at least 1 deaf child diagnosed with non-syndromic deafness. In Vitro Fertilization (IVF) was performed, followed by PGD for a cousin couple with a 50% chance of having an affected child. Bi-allelic pathogenic mutations were found in a total of 12 families (~17 %); of which a couple was a PGD volunteer. The deaf woman in this family was homozygous and her husband was a carrier of the IVS1+1G>A gene mutation. Among 8 biopsied embryos, two healthy embryos were implanted which resulted in a single pregnancy and subsequent birth of a healthy baby boy. This is the first report of a successful application of PGD for hearing loss in Iran. Having a baby with a severe hearing impairment often imposes families with long-term disease burden and heavy therapy costs. Today PGD has provided an opportunity for high-risk individuals to avoid the birth of a deaf child.

Background Preimplantation genetic diagnosis (PGD) has been developed to detect genetic disorders before pregnancy which is usually done on blastomeres biopsied from 8-cell stage embryos obtained from in vitro fertilization method (IVF). Here we report molecular PGD results for diagnosing of beta thalassemia (beta-thal) which are usually accompanied with evaluating chromosomal aneuploidies, HLA typing and sex selection. Methods In this study, haplotype analysis was performed using short tandem repeats (STRs) in a multiplex nested PCR and the causative mutation was detected by Sanger sequencing. Results We have performed PGDs on 350 blastomeres from 55 carrier couples; 142 blastomeres for beta-thal only, 75 for beta-thal and HLA typing, 76 for beta-thal in combination with sex selection, and 57 for beta-thal and aneuploidy screening. 150 blastomeres were transferable, 15 pregnancies were happened, and 11 babies born. We used 6 markers for beta-thal, 36 for aneuploidy screening, 32 for sex selection, and 35 for HLA typing. To our knowledge combining all these markers together and the number of STR markers are much more than any other studies which have ever done. Conclusions PGD is a powerful diagnostic tool for carrier couples who desire to have a healthy child and wish to avoid medical abortion.

Background: Preimplantation genetic diagnosis (PGD) has been developed to detect genetic disorders before pregnancy which is usually done on blastomeres biopsied from 8-cell stage embryos obtained from in vitro fertilization method (IVF).Here we report molecular PGD results for diagnosing of beta thalassemia (beta-thal) which are usually accompanied with evaluating chromosomal aneuploidies, HLA typing and sex selection.Methods: In this study, haplotype analysis was performed using short tandem repeats (STRs) in a multiplex nested PCR and the causative mutation was detected by Sanger sequencing.Results: We have performed PGDs on 350 blastomeres from 55 carrier couples; 142 blastomeres for beta-thal only, 75 for beta-thal and HLA typing, 76 for beta-thal in combination with sex selection, and 57 for beta-thal and aneuploidy screening. 150 blastomeres were transferable, 15 pregnancies were happened, and 11 babies born.We used 6 markers for beta-thal, 36 for aneuploidy screening, 32 for sex selection, and 35 for HLA typing. To our knowledge combining all these markers together and the number of STR markers are much more than any other studies which have ever done.Conclusions: PGD is a powerful diagnostic tool for carrier couples who desire to have a healthy child and wish to avoid medical abortion.

Background: Pre-implantation Genetic Diagnosis (PGD) has recently been introduced as a reproductive choice for individuals who carry a disease-causing BRCA1/2 mutation. Since this technology has not yet been launched for patients at the Cancer Institute of Imam Khomeini Hospital harboring gene mutations that predispose patients to breast cancer, this study aimed to introduce a PGD-based model using a single cell lymphocyte instead of an embryonic blastomere. Methods: Two affected and unrelated women with a known mutation in BRCA1/2 were enrolled in this study. Each patient (together with her siblings) was considered as an embryo derived from a hypothetical couple. Blood samples were collected from these individuals as well as their parents. Linkage analysis was performed. Following this process, a mutation-free individual and a mutation carrier was selected from the first and second family, respectively. A single lymphocyte was then extracted from their freshly taken peripheral blood, and afterwards Nested Multiplex PCR was performed. Results: PGD confirmed that the individual from the first family is free of a mutation and the second one is a pathogenic mutation carrier. Conclusion: Our results suggested that PGD is a viable choice to offer to families with "Hereditary Breast Cancer Syndrome", who have been diagnosed with a known pathogenic mutation. Our introduced model can be used as a possible option by other laboratories that are planning to launch this technology.

Introduction Breast cancer is the most common cancer in women and the second most common cause of cancer mortality in women, after lung cancer (Siegel, Miller et al. 2015). BRCA1 and BRCA2 are major genes related to hereditary breast cancer and are inherited in autosomal dominant form. Finding the causative mutated gene is very important and can help in prenatal diagnosis (PND) and pre-implantation Genetic Diagnosis (PGD) for high risk families. Linkage studies with the help of STR markers are very helpful to track the possible mutated gene. These markers also help in detecting "Allele Drop out" (ADO) in PGD procedure. This study aimed to find polymorphic STR markers linked to the BRCA1 gene for use in linkage studies in extended pedigrees and also in of PND & PGD studies. Materials and Methods: 50 unrelated individuals were genotyped to assess the allele frequencies, heterozygosity of the selected markers. Polymorphic STR markers were selected from Tandem Repeats Finder and Sequence-based Estimation of Repeat Variability databases (3, 4) (Legendre, Pochet et al. 2007) websites (Benson 1999). Suitable primers were designed to be to set up a multiplex-PCR reaction. Genotyping of each individual were performed using fragment analysis by ABI Genetic Analyzer 3130.Statistical analysis was performed using GenAlEx6.03. Results: Our results showed that the heterozygosity of selected markers were between 41%-95%. Totally, 43 alleles were observed. The highest heterozygosity was observed for D17SD-BRCA17.41and the lowest for D17SDBRCA126. 04. 6, 3,5 ,7 ,3 ,10,9 alleles were seen for D17SUBRCA118.85, D17SUBRCA115.53, D17SUBRCA113.07, D17SUBRCA18.06, D17SUBRCA11.89, D17SDBRCA17.41, D17SDBRCA126.04 respectively. Conclusions: Using these markers in a multiplex-PCR can be a simple, cheap, and fast tool for finding the possible mutated gene for cancer in the families under investigation. The next step would be direct sequencing of candidate gene. It is also very helpful for finding carrier members of the family.