Rapid diagnosis of α0 -thalassemia using the relative quantitative PCR and the dissociation curve analysis
| Autor: | P. Charoenporn, S. Sangkitporn, B. Songkharm, K. Wangkahat, S. K. Sangkitporn, A. Sangnoi |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
congenital
hereditary and neonatal diseases and abnormalities Thalassemia Alpha (ethology) Oxygen–haemoglobin dissociation curve Hematology Biology medicine.disease Molecular biology law.invention chemistry.chemical_compound Real-time polymerase chain reaction chemistry law hemic and lymphatic diseases Genotype medicine SYBR Green I Quantitative analysis (chemistry) Polymerase chain reaction |
| Zdroj: | Clinical & Laboratory Haematology. 25:359-365 |
| ISSN: | 0141-9854 |
| DOI: | 10.1046/j.0141-9854.2003.00549.x |
| Popis: | A method for diagnosis of alpha(o)-thalassemia was developed based on detection of accumulated PCR product using SYBR Green I, a double-stranded DNA binding dye, and a fluorescence-detecting thermocycler. Primers were designed to specifically amplify - -SEA and - -Thai deletions of alpha(o)-thalassemia. Albumin was selected as the reference gene. The comparative CT method was used to quantitate the target gene relative to the albumin gene. Dissociation curve analysis was used as a qualitative tool to distinguish different types of alpha(o)-thalassemia. In this study, the melting temperatures of the - -Thai and - -SEA deletions were 83 and 86 degrees C, respectively. Application of the assay for the diagnosis of alpha(o)-thalassemia heterozygotes and homozygotes is reported. The assay is highly robust and amenable to high throughput. It is thus a useful tool for the rapid detection of alpha(o)-thalassemia in populations with a high frequency of alpha(o)-thalassemia, - -SEA and - -Thai deletions. |
| Databáze: | OpenAIRE |
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