Development of the rapid diagnosis method of hemoglobinopathies, thalssemias and abnormal hemoglobins.

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Blood samples from patients expected to have hemoglobinopathy from the whole area of Japan and from transfusion dependent patients from Myanmar with severe anemia were analysed for the purpose of development of the rapid diagnosis method of hemoglobinopathies by the technique of molecular engineering.To the patients expected to p-thalassemia from Hb analysis, the ARMS (Amplification Refractory MutationSystem) method is useful for the detection of 5 mutations found commonly in Japan (-31CapA -> G, CD41/42TTCTTT→TT, CD90GAG→TAG, IVS II-1G→A, IVS II-654C→T) and those found in Myanmar (CD17 →T, IVS I-1G→T, IVS I-5G→C, CD41/42TTCTTT→TT, IVS II-654C→T), being discovered in about 70 percentof patients of both countries. However, this is difficult to adopt as a screening method, it is necessary to set up areaction condition to detect every mutation at the same ARMS. Additionally, the detection of an unknown or raremutation is required the PCR/direct sequencing.Major of abnormal Hb found in Myanmar is Hb E, that is easily found by agarose gel electrophoresis of PCRproduct digested with Mnl I. In the present study, the carriers of rare Hbs of Hb S and Hb Monroe had been foundIn Japan where found the various kind of Hb variants the PCR-sequencing method is necessary for the diagnosis.On the other hand, a-thalassemia mutations found in both countries were only two mutations of -α3.7 and - -SEA, and the mutations of -α4.2, - -MED, - -Fil and - -Thai were not detected. It is considered that the detection ofa-thalassemia mutations must be enough to take a technique of multiplex PCR-agarose gel electrophoresis.