使用自動化儀器Trinity Biotech Ultra II 和Sebia MINICAP 偵測高度不穩定血紅素蛋白：Hb Madrid

Use of Automated Trinity Biotech Ultra II and Sebia MINICAP to Detect a Severely Unstable Hemoglobin Variant: Hb Mardrid

我們利用DNA定序確定了一個台灣人β-globin gene codon 115 GCC-CCC（Ala> Pro）的突變。此變異血色素名為Hb Madrid，該β-chain突變造成了血紅素的嚴重不穩定。大多數高不穩定性血紅素蛋白（Hyperunstable hemoglobin variants；HUH）幾乎都無形成穩定的蛋白質，無法使用偵測蛋白質平台測出，包括質譜儀。此血紅素的變異無法使用Bio-RAD D10 HPLC平台檢測，但確可以透過陽離子交換HPLC（Trinity Biotech Ultra II）和Sebia Minicap發現。此β-globin gene在heterozygotes突變情況下呈現顯性β-海洋性貧血，存在嚴重的溶血性貧血、reticulocyte增多、在骨髓具有明顯的紅血球細胞增生、脾腫大，並且Hb F顯著增加。 We have identified through sequencing of amplified DNA a GCC-CCC(Ala>Pro), mutation incodon 115 of the β-globin gene in Taiwanese. It is called Hb Mardrid, is a Hyperunstablehemoglobin variants (HUH). Most Hyperunstable hemoglobin variants (HUH) result frommutations that are transcribed and translated but have a defect so severe that they do not form aprotein that is detectable by any method, including mass spectrometry and stability studies. Thisβ-chain is severely unstable and could not be identified as hemoglobin variant by high performanceliquid chromatography (Bio-RAD D10) methods. But It could be identified by Cation-ExchangeHPLC (Trinity Biotech Ultra II) and Sebia Minicap. It is presence results in a dominant type ofβ-thalassemia in the two heterozygotes, with severe hemolytic anemia, reticulocytosis, hypercellularmarrow with prominent erythroid hyperplasia, splenomegaly, and had a marked increase in fetalhemoglobin synthesis.

We have identified through sequencing of amplified DNA a GCC-CCC(Ala>Pro), mutation incodon 115 of the β-globin gene in Taiwanese. It is called Hb Mardrid, is a Hyperunstablehemoglobin variants (HUH). Most Hyperunstable hemoglobin variants (HUH) result frommutations that are transcribed and translated but have a defect so severe that they do not form aprotein that is detectable by any method, including mass spectrometry and stability studies. Thisβ-chain is severely unstable and could not be identified as hemoglobin variant by high performanceliquid chromatography (Bio-RAD D10) methods. But It could be identified by Cation-ExchangeHPLC (Trinity Biotech Ultra II) and Sebia Minicap. It is presence results in a dominant type ofβ-thalassemia in the two heterozygotes, with severe hemolytic anemia, reticulocytosis, hypercellularmarrow with prominent erythroid hyperplasia, splenomegaly, and had a marked increase in fetalhemoglobin synthesis.