<正>Objective :To evaluate the possibility of the technology involving PEP and RDB for detectingβ-thalassaemia multipoint mutations from a single cell simultaneously. Methods: A set of allele specific oligonucleotide （ASO） probes used for detecting 8 familiarβ-thalassaemia mutations （CD41-42. IVS-Ⅱ-654

CD17

TATA box nt-28

CD71-72

TATA box nt-29

CD26

IVS-Ⅰ-5） were immobilized on a strip of nylon membrane. The genome of a individual cell was amplified by primer extension preamplification （PEP） with the mixture of 15-base random oligonucleotides. The aliquots from PEP were used to amplify the objective gene fractions ofβ-thalassaemia gene by nested or semi-nested PCR. The membrane was hybridized with the final amplified products and then treated with Streptavidin-HRP and color development. Results:Totally 30 lymphocytes were picked up from blood samples of 1 healthy female and 4 patients with knownβ-thalassaemia mutations respectively. Each single lymphocyte was lysed in the proteinase K buffer. The amplification efficacy was 94. 0% and alle drop-out （ADO） rate was 8. 0%. Revert dot blot （RDB） was applied to the final amplified products from the 5 participants. The results of diagnosis were the same to the expected

and their genotypes were N/N

CD17（A→T）/N

IVS-Ⅱ-654（C→T）/CD17（A→T）

CD41-42（-CTTT）/N and TATA box nt-28（A→G）/N

respectively. Conclusion: The technology involving PEP and RDB could detect multipleβ-thalassaemia mutations from a single cell simultaneously

and the research provides experimental evidences for the feasibility of applying PEP and DNA array technology to screening multiple genetic mutations from a single cell

and will be applied to preimplantation genetic diagnosis and non-invasive prenatal diagnosis forβ-thalassaemia.