Differential Detection of α or β-Globin Gene Deletion by Semi-Quantitative Triplex PCR

Prevention of β-thal is an ongoing program in Iran. There are a large number of couples who have been suspicious in α or β-thal. Exact determination being carrier of α-thal or β-thal at the molecular level is crucial for counseling and prenatal diagnosis. In order to determine the presence or absence of deletion in β- , α1- , or α2- globin genes, a triplex PCR is designed which quantitatively determines presence or absence of deletion in either of the above genes. PCR was optimized testing different conditions. Crucial parameters were found to be DNA extraction method (salting out was best), dNTP concentration, and presence of DMSO and glycerol. The study have been done on 10 normal and 50 individuals referred to us as being possible carrier of α or β. PCR products with intact (no deletion) gene showed band densities different from persons either having β-gene deleted or either or both α-genes deleted. If no deletion is detected, the sample can be a subject for point mutation study. Several repeated tests with samples known to have deletion in β- or α-globin genes proved our quantitative method being very accurate and useful.

کلیدواژه ها:

thalassemia ، deletion ، alpha ، beta ، PCR