OBJECTIVETo establish a rapid method for detection of alpha-globin gene αααbased on droplet digital PCR (ddPCR) technique.

METHODSThe differential sequence between the X1 and Y1 box of α1 gene was selected as the amplicon of the target gene with β-actin as the reference gene. The specific primers and TaqMan probes were designed, and then a quantitative method for detecting the copy number was established based on ddPCR technique. The sensitivity and accuracy of the method were evaluated by detecting 28 samples of known genotypes and 60 clinical samples.

RESULTSThe ddPCR-based method accurately identified the genotypes of all the 28 samples with known genotypes and detected 5 cases of αα/αααfrom the 60 clinical samples, and the results were verified by MLPA. The sensitivity and accuracy of this method were both 100% for detecting alpha-globin gene ααα.

CONCLUSIONThis ddPCR-based method for detecting αααtriplet can be applied for population screening and in routine clinical molecular diagnosis with simple operation, rapid analysis and accurate results.