Single-tube Multiplex Polymerase Chain Reaction For Detection Of ¦Á-thalassemia Genotype

Objective To develop techniques based on single-tube multiplex polymerase chain reaction (PCR) for diagnosis of common a- thalassemia (a-thal) genotypes in china. Methods 146 Hb H disease samples and 74 fatel DNA from chorionic villi samples (CVS) or amniocyte samples were detected by the two sets of single-tube multiplex polymerase chain reaction (PCR) and classic PCR methods which had been commonly used respectively. Results The single-tube PCR can detect SEA α-thal-1, -α3.7 α-thal-2, -α4.2 a-thal-2, Hb Constant Spring (Hb CS) and Hb Quong Sze (Hb QS) genotypes, and the results were the same as those detected by classic PCR methods. Among 146 Hb H disease patients, 83 cases were riondeletional Hb H disease , accounting for 56.9% (81 cases of -SEA/αcsα, 1 case of --SEA/αQSα, 1 case of -SEA/αTα unknown mutation); 63 cases were deletional Hb H disease, accounting for 43.2% ( 41 cases of -SEA/-α 3.7, 22 cases of -SEA/-α4.2) Among 74 fatel DNA samples, 40 cases were normal (αα/αα), while a-thal-1 heterozygotes 25, Hb Bart's hydrops fetalis syndrome 4, deletional Hb H disease 4(3 for -SEA/-α4.2, 1 for -SEA /-α3.7) and Hb CS heterozygotes 1. Conclusion The single-tube multiplex PCR for detecting a-thal genotypes especially Hb H disease was accurate and sensitive. It was simple and easy to be practice for screening a-thal genotypes.