A novel deletion of  -globin promoter causing high HbA2 in an Indian population

Identification of rare and novel deletions that cause (δβ)0‐thalassaemia and hereditary persistence of foetal haemoglobin in Indian population

“…Copy numbers of γ‐, δ‐ and β‐globin genes were measured by a gene dosage quantitative multiplex fluorescent PCR‐PCR (GD‐QF‐MPCR) as we described recently . We used Hot Star Master Mix (Qiagen, GmbH, Hilden, Germany) with 0.5 μ m primers for 20–22 cycles with 200 ng of DNA, and the fluorescently labelled amplified products were analysed by capillary electrophoresis using ABI genetic analyser and the peak heights were determined by GeneMapper 4.0 (Applied Biosystems).…”

“…Using this method, the breakpoints of the deletions cannot be located as the probes that bind to the genomic locus are widely spaced and do not span the whole genomic region where these deletions occur. We developed quantitative fluorescent multiplex PCR (QF‐MPCR), which are cheaper and easier to perform, to detect the large deletions in the β‐globin cluster . This method could also facilitate locating the breakpoints successfully.…”

Rapid molecular identification of a rare β-globin gene deletion & its clinical implication

“…A combined strategy of MLPA, QF-PCR and gap-PCR was thus used to detect this deletion. Previously, the 4056 bp deletion was first identified by Mayuranathan et al 6 , 2012, in a patient of Gujarat origin. They used combined MLPA and PCR techniques to identify this deletion wherein the PCR generated only one deletion-specific band of 1.1 kbp.…”

“…Net software (Coffalyser.Net - MRC, Holland). Further, a simplified single tube gap-PCR was performed with the help of previously reported primers sequences 6 . The gap-PCR programme employed to detect the deletion was as follows: initial denaturation at 94°C for five minutes, followed by 30 cycles of denaturation: 94°C for one minute, annealing at 58°C for one minute, extension at 72°C for one minute and a final extension at 72°C for five minutes.…”

“…Large deletions in the β-globin cluster escape traditional diagnostic techniques 6 . In our study as well, the CRDB, ARMS PCR and direct DNA sequencing failed to identify the presence of 4056 bp deletion.…”