Development of the simple-rapid diagnosis method of hemoglobinopathies, thalassemias and abnormal hemoglobins

1.应用聚合酶链式反应-直接测序法对两例新发现的异常Hb福井[α139(Hc1)Lys→Asn]和Hb Turriff[α99(G6)Lys→Glu]的DNA进行了直接测序分析，发现了与Hb Hanamaki和Hb Tokoname具有相同密码子的a 2基因突变，并对3种单倍型进行了检测。3.测定250名泰国受试者的CBC后，用HLC-723G7全自动Hb A_2分析仪测定Hb A_2水平。β-Thal采用ARMS和聚合酶链式反应直接测序法，α-Thal采用GAP法。从这些数据得到的HbA_2-MCV和HbA_2-Mch的散点图谱表明，βThal携带者HbA_2水平高于4%，Mcv和Mch低于正常，而αThal携带者HbA_2、Mcv和Mch水平低于正常，其中αThal-1病例比α-Thal-2病例严重。4.将快照方法应用于HbA和Thal异常的检测，显示了一次检测多种突变的可能性。此外，实时定量聚合酶链式反应方法的应用有助于诊断长区缺失的β-Thal。

The diagnosis and the simple-rapid diagnostic method for the abnormal hemoglobins(Hb) and thalassemias(thal) were discussed.1.PCR-direct sequence analysis of DNAs from two Japanese identified to be a new abnormal Hb Fukui[α 139(HC1)Lys→Asn] with the mutation in the a 2 gene which is the same codon to Hb Hanamaki and Hb Tokoname, and Hb Turriff[α 99(G6)Lys→Glu] with the mutation in the α 1 gene.2.The Myanmar β^E-gene was identified by PCR-RFLP, and among three haplotypes detected, the C type + --+ + + was characteristic for Myanmar.3.After the CBC of 250 Thai subjects was determined, the Hb A_2 level was determined by an automated Hb A_2 analyzer, HLC-723G7. β-Thal was diagnosed by the ARMS and PCR-direct sequence methods, and α-thal done by the Gap method. The scattered grams of Hb A_2-MCV and Hb A_2-MCH resulting from these data, suggested that the β thal carriers have higher Hb A_2 level than 4% and lower MCV and MCH than the normal, and the α thal carriers lower level of Hb A_2, MCV and MCH than the normal, in which the α thal-1 cases were rather severe than the α-thal-2 cases.4.The SNaPshot method was applied to the detection of abnormal Hbs and thals, and showed the possibility to find out many mutations at one reaction. Additionally, the application of the Real Time PCR method was useful for the diagnosis of β thal with the deletion of long region.