Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study
<p>Abstract</p> <p>Background</p> <p>Hereditary Hemochromatosis (HH) is an autosomal recessive disorder highlighted byiron-overload. Two popular mutations in <it>HFE</it>, p.C282Y and p.H63D, have been discovered and found to associate with HH in different e...
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| Format: | Article |
| Language: | English |
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BMC
2006-05-01
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| Series: | BMC Medical Genetics |
| Online Access: | http://www.biomedcentral.com/1471-2350/7/43 |
| _version_ | 1798029676480823296 |
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| author | Al-Hamed Mohamed Al-Kayal Fadi Alsmadi Osama A Meyer Brian F |
| author_facet | Al-Hamed Mohamed Al-Kayal Fadi Alsmadi Osama A Meyer Brian F |
| author_sort | Al-Hamed Mohamed |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Hereditary Hemochromatosis (HH) is an autosomal recessive disorder highlighted byiron-overload. Two popular mutations in <it>HFE</it>, p.C282Y and p.H63D, have been discovered and found to associate with HH in different ethnic backgrounds. p.C282Y and p.H63D diagnosis is usually made byrestriction enzyme analysis. However, the use of this technique is largelylimited to research laboratories because they are relativelyexpensive, time-consuming, and difficult to transform into a high throughput format.</p> <p>Methods</p> <p>Single nucleotide variations in target DNA sequences can be readily identified using molecular beacon fluorescent probes. These are quenched probes with loop and hairpin structure, and they become fluorescent upon specific target recognition. We developed high throughput homogeneous real-time PCR assays using molecular beacon technology, to genotype p.C282Y and p.H63D variants. Representative samples of different genotypes for these variants were assayed by restriction enzyme analysis and direct sequencing as bench mark methods for comparison with the newly developed molecular beacon-based real-time PCR assay.</p> <p>Results</p> <p>Complete concordance was achieved by all three assay formats. Homozygotes (mutant and wildtype) and heterozygotes were readily differentiated by the allele specific molecular beacons as reported by the associated fluorophore in the real-time assay developed in this study. Additionally, these assays were used in a high throughput format to establish the allele frequency of C282Y and H63D in Saudis for the first time.</p> <p>Conclusion</p> <p>These assays may be reliably applied as a diagnostic test or large scale method for population screening.</p> |
| first_indexed | 2024-04-11T19:29:01Z |
| format | Article |
| id | doaj.art-d9fe1ea3d39c48a6b70cf2b24ac3ae68 |
| institution | Directory Open Access Journal |
| issn | 1471-2350 |
| language | English |
| last_indexed | 2024-04-11T19:29:01Z |
| publishDate | 2006-05-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Medical Genetics |
| spelling | doaj.art-d9fe1ea3d39c48a6b70cf2b24ac3ae682022-12-22T04:07:03ZengBMCBMC Medical Genetics1471-23502006-05-01714310.1186/1471-2350-7-43Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based studyAl-Hamed MohamedAl-Kayal FadiAlsmadi Osama AMeyer Brian F<p>Abstract</p> <p>Background</p> <p>Hereditary Hemochromatosis (HH) is an autosomal recessive disorder highlighted byiron-overload. Two popular mutations in <it>HFE</it>, p.C282Y and p.H63D, have been discovered and found to associate with HH in different ethnic backgrounds. p.C282Y and p.H63D diagnosis is usually made byrestriction enzyme analysis. However, the use of this technique is largelylimited to research laboratories because they are relativelyexpensive, time-consuming, and difficult to transform into a high throughput format.</p> <p>Methods</p> <p>Single nucleotide variations in target DNA sequences can be readily identified using molecular beacon fluorescent probes. These are quenched probes with loop and hairpin structure, and they become fluorescent upon specific target recognition. We developed high throughput homogeneous real-time PCR assays using molecular beacon technology, to genotype p.C282Y and p.H63D variants. Representative samples of different genotypes for these variants were assayed by restriction enzyme analysis and direct sequencing as bench mark methods for comparison with the newly developed molecular beacon-based real-time PCR assay.</p> <p>Results</p> <p>Complete concordance was achieved by all three assay formats. Homozygotes (mutant and wildtype) and heterozygotes were readily differentiated by the allele specific molecular beacons as reported by the associated fluorophore in the real-time assay developed in this study. Additionally, these assays were used in a high throughput format to establish the allele frequency of C282Y and H63D in Saudis for the first time.</p> <p>Conclusion</p> <p>These assays may be reliably applied as a diagnostic test or large scale method for population screening.</p>http://www.biomedcentral.com/1471-2350/7/43 |
| spellingShingle | Al-Hamed Mohamed Al-Kayal Fadi Alsmadi Osama A Meyer Brian F Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study BMC Medical Genetics |
| title | Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study |
| title_full | Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study |
| title_fullStr | Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study |
| title_full_unstemmed | Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study |
| title_short | Frequency of common <it>HFE </it>variants in the Saudi population: a high throughput molecular beacon-based study |
| title_sort | frequency of common it hfe it variants in the saudi population a high throughput molecular beacon based study |
| url | http://www.biomedcentral.com/1471-2350/7/43 |
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