ABSTRACT
This study was done with a view to find a correlation between two molecular tests for beta Thalassemia, our in-house developed haemoglobin DNA mutation analysis using ARMS PCR and a Commercial Line Probe Assay. De-identified samples from known beta thalassemia patients characterised by HPLC for HbA2, Peripheral smear [Target cells] and CBC [microcytosis and erythrocytosis] were used for the study. DNA was extracted using the DTAB/CTAB method. Amplified DNA from the samples was hybridised for mutations using a line probe assay. The extracted DNA was also examined for wild type genes and mutant genes using an Amplification refractory mutation system [ARMS] PCR. In this study fifteen beta Thalassemia patients were involved. The in-house ARMS PCR tested for six mutations and detected thalassemia trait in 66.7% of the samples tested for. The line probe assay tested for 22 mutations and detected thalassemia trait in 93.7% of cases examined. One case was missed by both methods and will require sequencing. The importance of stratification of testing for a cost effective strategy for Thalassemia diagnostics is discussed
ABSTRACT
This pilot study was initiated with a view to find alpha thalassemia genotypes on de-identified samples from patients diagnosed with anaemia at the Centre for Advanced Biomedical research and Innovation [CABRI] at Gulf Medical University [GMU] in June 2015. Amplified DNA from the samples was probed for mutations using a line probe assay. Results obtained are presented. The study has shown the 3.7 single gene deletion in three cases, and alpha 2 IVS1 [-5nt] mutation seen in one case suggesting these cases have alpha + thalassemia. One sample showed wild type alpha 2 Poly A missing along with the alpha 2 poly a-1 [AATAAA>AATAAG] mutation with a suggestive diagnosis of HbH disease. A SEA double gene deletion was seen in one case suggesting alpha 0-thalassemia. Further studies are being carried out to enhance the data base