Application of the Single-Molecule Real-Time Technology (SMRT) for Identification of HKαα Thalassemia Allele

Objective: Single-molecule real-time technology (SMRT) is a sequencing technology using the DNA polymerases and fluorescently tagged nucleotides to accurately sequence DNA strands. The purpose of this study was to evaluate the detection accuracy of SMRT for identification of the Hong Kong alpha alpha (HK alpha alpha) thalassemia allele. Methods: We conducted a blinded study of 33 samples of known HK alpha alpha alleles. These alleles were detected using SMRT to evaluate accuracy. Results: We conducted a blinded study of 33 known HK alpha alpha samples and found all HK alpha alpha. variants detected by SMRT to be concordant with those independently assigned by gap-polymerase chain reaction (gap-PCR), reverse dot blot hybridization, and 2-round nested PCR. In addition, SMRT detected 2 beta-thalassemia variants that were missed by conventional techniques. Conclusion: The results demonstrate that SMRT offers a higher detection accuracy of thalassemia rare and new loci. It is an efficient, reliable, and broad-spectrum test that can be widely used for thalassemia screening in the clinic.

Automated summary

This study validated the detection accuracy of Single-molecule real-time technology (SMRT) for thalassemia alleles and found it to be superior to conventional techniques, especially in detecting rare and new variants. Therefore, SMRT can be widely used as an efficient, reliable, and broad-spectrum method for thalassemia screening.