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

用分子工程技术对来自日本各地的疑似血色素沉着症患者和来自缅甸的严重贫血输血依赖患者的血液样本进行分析，以建立血红蛋白病的快速诊断方法。对于从Hb分析中预计患有β地中海贫血的患者，ARMS(Amplify Refired MutationSystem)方法可用于检测在日本常见的5种突变(-31CapA-&GT；CD41/42TTCTTT→TT、CD90GAG→Tag、IVS II-1G→A、IVS II-654C→T)和在缅甸发现的(CD17→T、IVS I-1G→T、IVS I-5G→C、CD41/42TTCTTT→TT、IVS II-654C→T)，在两国约70%的患者中发现。然而，这很难作为一种筛查方法采用，有必要设置反应条件来检测同一臂上的每一种突变。在缅甸发现的异常Hb主要是Hb E，用MNL I酶切的PCR产物琼脂糖凝胶电泳法很容易发现Hb E。在本研究中，在日本发现了罕见的Hb携带者S和Hb Monroe，在日本发现了各种Hb变异体，需要用聚合酶链式反应测序方法进行诊断。另一方面，在两国发现的α-地中海贫血突变只有-α3.7和-SEA两个突变，-α4.2，-MED，--FIL和--泰语未被检测到。认为采用多重聚合酶链式反应-琼脂糖凝胶电泳法对α-地中海贫血基因突变的检测是必要的。

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.