Genotyping of ß-globin gene mutations in single lymphocytes: A preliminary study for preimplantation genetic diagnosis of monogenic disorders

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dc.contributor.authorDurmaz B.
dc.contributor.authorÖzkınay F.
dc.contributor.authorOnay H.
dc.contributor.authorKaraca E.
dc.contributor.authorAydinok Y.
dc.contributor.authorTavmergen E.
dc.contributor.authorVrettou C.
dc.contributor.authorTraeger-Synodinos J.
dc.contributor.authorKanavakis E.
dc.date.accessioned2019-10-27T08:33:04Z
dc.date.available2019-10-27T08:33:04Z
dc.date.issued2012
dc.departmentEge Üniversitesien_US
dc.description.abstractHemoglobinopathies, especially ß-thalassemia (ß-thal), represent an important health burden in Mediterranean countries like Turkey. Some couples prefer the option of preimplantation genetic diagnosis (PGD). However, clinical application of PGD, especially for the monogenic disorders is technically demanding. To ensure reliable results, protocols need to be robust and well standardized. Ideally PGD-PCR (polymerase chain reaction) protocols should be based on multiplex and fluorescent PCR for analysis of the disease-causing mutation(s) along with linked markers across the disease-associated locus. In this study, we aimed to constitute a protocol in single cells involving first round multiplex PCR with primers to amplify the region of the ß-globin gene containing the most common mutations. Two microsatellites linked to the ß-globin gene cluster (D11S4891, D11S2362) and two unlinked (D13S314, GABRB3) microsatellite markers, were used to rule out allele dropout (ADO) and contamination; followed by nested real-time PCR for genotyping the ß-globin mutations. We also investigated the allele frequencies and heterozygote rates of these microsatellites in the Turkish population that have not been reported to date. This protocol was tested in 100 single lymphocytes from heterozygotes with known ß-globin mutations. Amplification failure was detected in one lymphocyte (1%) and ADO was observed in two lymphocytes (2%). No contamination was detected. All results were concordant with the genotypes of the patients. Overall, this protocol was demonstrated to be sensitive, accurate, reliable and rapid for the detection of ß-globin mutations in single cells and shows potential for the clinical application of PGD for hemoglobinopathies in the Turkish population. Copyright © Informa Healthcare USA, Inc.en_US
dc.identifier.doi10.3109/03630269.2012.675891en_US
dc.identifier.endpage243en_US
dc.identifier.issn0363-0269
dc.identifier.issue3en_US
dc.identifier.pmid22524255en_US
dc.identifier.scopusqualityQ3en_US
dc.identifier.startpage230en_US
dc.identifier.urihttps://doi.org/10.3109/03630269.2012.675891
dc.identifier.urihttps://hdl.handle.net/11454/26642
dc.identifier.volume36en_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.relation.ispartofHemoglobinen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectß-Globin geneen_US
dc.subjectReal-time polymerase chain reaction (PCR)en_US
dc.subjectSingle cell polymerase chain reaction (PCR)en_US
dc.subjectThalassemiaen_US
dc.titleGenotyping of ß-globin gene mutations in single lymphocytes: A preliminary study for preimplantation genetic diagnosis of monogenic disordersen_US
dc.typeArticleen_US

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