Molecular genetics of beta thalassaemia in Asian Indians
<p>The primary aim of this thesis was to outline an approach for the prenatal diagnosis of β-thalassaemia in the Asian Indian population by DNA analysis. A polymerase chain reaction (PCR) based, nonradioactive and rapid technique, allele specific PCR, was successfully developed for the detecti...
| Autores principales: | , |
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| Formato: | Tesis |
| Lenguaje: | English |
| Publicado: |
1992
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| _version_ | 1826304849249042432 |
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| author | Varawalla, N Varawalla, Nermeen Y |
| author2 | Old, J |
| author_facet | Old, J Varawalla, N Varawalla, Nermeen Y |
| author_sort | Varawalla, N |
| collection | OXFORD |
| description | <p>The primary aim of this thesis was to outline an approach for the prenatal diagnosis of β-thalassaemia in the Asian Indian population by DNA analysis. A polymerase chain reaction (PCR) based, nonradioactive and rapid technique, allele specific PCR, was successfully developed for the detection of β-thalassaemia mutations. A large sample of 656 unrelated carriers from seven different regions of the Indian subcontinent was studied by allele specific PCR and DNA sequence analysis. Sixteen different β-thalassaemia mutations were identified, two of which were new mutations. Of these five common mutations accounted for 91.7% of β-thalassaemia alleles.</p><p>The β-globin gene haplotypes of 419 β-Th and 196 β - A chromosomes were constructed. On analysis of which it was inferred that β-thalassaemia mutations occurred relatively recently on existing chromosomal backgrounds and then they experienced positive selection. A strong but not invariant haplotype-mutation linkage was observed. A regional variation in the distribution of β-thalassaemia mutations was found.</p><p>α-Globin gene mapping studies identifed the single α-globin gene deletion in 24 out of 51 unrelated Asian Indians who were suspected to have α-thalassaemia. It is likely that the remaining carriers have nondeletional α-thalassaemia determinants.</p><p>To perform preimplantation diagnosis of β-thalassaemia, by analysis of a 10-30 cell embryonic biopsy, a PCR protocol was developed. Using two rounds of PCR with nested primers, successful amplification of a 597 bp fragment of the β-globin gene was achieved from as few as two embryonic cells. The problem of false positive amplification was encountered which appeared to be resolved by UV transillumination of the pre-amplification PCR mix. By allele specific PCR with nested primers it was possible to identify the presence or absence of five β-thalassaemia mutations from 10 pg of template DNA (equivalent to approximately two diploid cells).</p><p>Thalassaemia control in India is a complex issue; the financial, social and demographic factors involved were considered and recommendations made.</p> |
| first_indexed | 2024-03-07T06:23:59Z |
| format | Thesis |
| id | oxford-uuid:f3a2a0a7-3d14-4dcf-a6fc-616db75119bf |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:23:59Z |
| publishDate | 1992 |
| record_format | dspace |
| spelling | oxford-uuid:f3a2a0a7-3d14-4dcf-a6fc-616db75119bf2022-03-27T12:13:40ZMolecular genetics of beta thalassaemia in Asian IndiansThesishttp://purl.org/coar/resource_type/c_db06uuid:f3a2a0a7-3d14-4dcf-a6fc-616db75119bfMolecular aspectsDiagnosisMolecular geneticsThalassemiaEnglishPolonsky Theses Digitisation Project1992Varawalla, NVarawalla, Nermeen YOld, JWeatherall, DOld, JWeatherall, D<p>The primary aim of this thesis was to outline an approach for the prenatal diagnosis of β-thalassaemia in the Asian Indian population by DNA analysis. A polymerase chain reaction (PCR) based, nonradioactive and rapid technique, allele specific PCR, was successfully developed for the detection of β-thalassaemia mutations. A large sample of 656 unrelated carriers from seven different regions of the Indian subcontinent was studied by allele specific PCR and DNA sequence analysis. Sixteen different β-thalassaemia mutations were identified, two of which were new mutations. Of these five common mutations accounted for 91.7% of β-thalassaemia alleles.</p><p>The β-globin gene haplotypes of 419 β-Th and 196 β - A chromosomes were constructed. On analysis of which it was inferred that β-thalassaemia mutations occurred relatively recently on existing chromosomal backgrounds and then they experienced positive selection. A strong but not invariant haplotype-mutation linkage was observed. A regional variation in the distribution of β-thalassaemia mutations was found.</p><p>α-Globin gene mapping studies identifed the single α-globin gene deletion in 24 out of 51 unrelated Asian Indians who were suspected to have α-thalassaemia. It is likely that the remaining carriers have nondeletional α-thalassaemia determinants.</p><p>To perform preimplantation diagnosis of β-thalassaemia, by analysis of a 10-30 cell embryonic biopsy, a PCR protocol was developed. Using two rounds of PCR with nested primers, successful amplification of a 597 bp fragment of the β-globin gene was achieved from as few as two embryonic cells. The problem of false positive amplification was encountered which appeared to be resolved by UV transillumination of the pre-amplification PCR mix. By allele specific PCR with nested primers it was possible to identify the presence or absence of five β-thalassaemia mutations from 10 pg of template DNA (equivalent to approximately two diploid cells).</p><p>Thalassaemia control in India is a complex issue; the financial, social and demographic factors involved were considered and recommendations made.</p> |
| spellingShingle | Molecular aspects Diagnosis Molecular genetics Thalassemia Varawalla, N Varawalla, Nermeen Y Molecular genetics of beta thalassaemia in Asian Indians |
| title | Molecular genetics of beta thalassaemia in Asian Indians |
| title_full | Molecular genetics of beta thalassaemia in Asian Indians |
| title_fullStr | Molecular genetics of beta thalassaemia in Asian Indians |
| title_full_unstemmed | Molecular genetics of beta thalassaemia in Asian Indians |
| title_short | Molecular genetics of beta thalassaemia in Asian Indians |
| title_sort | molecular genetics of beta thalassaemia in asian indians |
| topic | Molecular aspects Diagnosis Molecular genetics Thalassemia |
| work_keys_str_mv | AT varawallan moleculargeneticsofbetathalassaemiainasianindians AT varawallanermeeny moleculargeneticsofbetathalassaemiainasianindians |