Real-Time PCR for the Rapid detection of common β-Thalassemia mutation in Gujarat

The inherited Haemoglobinopathis are autosomal recessive disorder including beta Thalassemia. Out of 200 different mutations causing β Thalassemia, IVS 1-1, IVS 1-5, Codon 41-42, Codon 8-9,and Codon 619 Bp deletion are most common in Gujarat Thalassemic patients. Rapid technique of prenatal diagnosis of only IVS 1-1, IVS 1-5 and Codon 8-9 have been standardized in our lab and Prenatal Diagnosis of these mutations was implemented. The Roche Light Cycler 480 (LC 480) system was evaluated for qualitative diagnosis of common mutations in β Thalassemia in Hb beta gene. A total number of 60 whole blood samples from carrier couples , 11 CVS and 9 Amniotic fluid samples were obtained from antenatal mothers .For testing; LC480 system is more suitable and versatile .Real time PCR platform in a routine laboratory settings for the diagnosis of common mutations in β Thalassemia as noted above is the basis of the present study.DNA Samples used for amplification where obtained from CVS or Amniotic fluid and whole Blood samples .The Standardized mutations in our lab were detected using 2 sets of probes and one common set of universal primer ,usually distinguishing mutants from the normal alleles. In Prenatal diagnosis of βThalassemia mutations accuracy and the speed are of paramount importance taking only one and half hours by the entire procedures in our present study.