| 摘要: | 有超過一千多篇血色素Hemoglobin（Hb）的變異型相關著作已被發表。血色素的疾病包括甲型地中海型貧血，乙型地中海貧血及其他變異血色素，是在台灣常見的遺傳性疾病。大多數的變異通常是α基因鏈或是β基因鏈上的基因突變。有鑑於此政府自從1993年開始推動孕婦海洋性貧血篩檢，此醫療政策使得重度β-thalassemia貧血患者的盛行率年平均也由1991至1996年的百萬分之52.3，降為1999至2003年的為百萬分之11.6。重型乙型海洋性貧血病患在台灣帶因率約5%～8%，而且這種遺傳的血液疾病是全世界分佈最廣泛、發生率最高，也最常見的單一基因遺傳疾病。由於相對較低的成本，分析血色素的醋酸纖維膜或瓊脂凝膠電泳仍然是應用於臨床醫院最普遍的方法，但是電泳的技術對於某些變異型的血紅素是無法檢測出來的。有鑑於此現今臨床上常利用的篩檢方法，包括血紅素電泳和高壓液相色層析法（HPLC）。這兩種方法，雖然被廣泛應用於許多臨床實驗室，但是對於變異血色素的檢驗都沒有絕對肯定的答案，除非再利用胺基酸定序或DNA確認試驗。
從1990年代開始蛋白質體檢驗的技術更加的純熟，質譜儀現今已被迅速應用到分析血色素的蛋白質。最近，電噴霧質譜electrospray ionization mass spectrometry (ESI-MS)和基質輔助雷射脫附游離飛行時間質譜儀matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)已經成為兩個主要在定性檢測變異型血色素的分析工具。本研究就是利用MALDI-TOF，其分析步驟簡單、結果又能夠準確分析到胺基酸的序列（該準確性等同分析到DNA序列），以及分析檢體需要量少等諸多優點，進行台灣常見變異血色素之研究。
結果顯示在我們分析的42例變異血色素的個案中有12例為Hb Phnom-Penh約占28.6%，8例為Hb E約占19.0%，10例為Hb J-meinung（J-Bangkok）約占23.8%，5例為Hb H約占11.9%其中3例另外含有
Hb Constant Spring（Hb CS），3例為Hb Owari約占7.1%，以及Hb G-Taichung（Q-Thailand）、Hb Prato、Hb Westmead、Hb Woodville各為1例各占2.4%，所有的個案利用HPLC、MALDI-TOF分析都能夠得到與DNA sequencing確認後相同的結果，綜合上述結果我們希望能夠藉由本研究建立起HPLC及MALDI-TOF在台灣常見的變異血色素的分析資料庫以作為未來臨床上篩檢變異血色素的另一種選擇。
Over more than one thousand paper about hemoglobin (Hb) variant has been published. Hemoglobin’s diseases, including alpha-thalassemia, beta-thalassemia and other variations, are common genetic diseases in Taiwan. Most genetic variation is usually α or β chain gene chain gene mutation. For this reason our government began promoting pregnant women since 1993, thalassemia, the health policy making severe beta- thalassemia the prevalence of anemia in patients also averaged from 1991 to 1996 was 52.3 per million, reduced from 1999 to 2003 of 11.6 ppm. Patients with severe beta-thalassemia carrier rate in Taiwan, about 5% to 8%, and this genetic blood disease in the world is most widely distributed, the highest incidence, but also the most common single gene genetic disease. As the relatively low cost, of hemoglobin in cellulose membrane or agarose gel electrophoresis is still the most widely used of clinical hospital, but electrophoresis technology for some variant of hemoglobin that can not be detected. In view of this current is often used in clinical screening methods, including hemoglobin electrophoresis and high pressure liquid chromatography (HPLC). These two methods, although being widely used in many clinical laboratories, but for confirm hemoglobin variants are no absolute definite answer that unless the use of amino acids sequencing or DNA sequencing confirm test.
From the beginning of the 1990s the technology of proteomics test more of the skilled, MS now has been quickly applied to the analysis of the protein hemoglobin. Recently, ESI-MS electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF) has been mainly in the qualitative detection of Hb variants analysis tools. Our study is the use of MALDI-TOF, the analytical procedure is simple, it can accurately analyze the amino acid sequence (equivalent to the accuracy of the analysis to the DNA sequence), and the analysis of samples need less, and many other advantages for Taiwan’s common hemoglobin variant study.
The results show that in our analysis of 42 cases of variant hemoglobin in these cases, 12 cases of Hb Phnom-Penh for about 28.6%, 8 cases of Hb E about 19.0%, 10 cases of Hb J-meinung (J-Bangkok) about 23.8%, 5 cases of Hb H accounted for about 11.9%, other 3 patients with Hb Constant Spring (Hb CS), 3 cases of Hb Owari about 7.1%, and Hb G-Taichung (Q-Thailand), Hb Prato, Hb Westmead, Hb Woodville 1 case each for each 2.4% of all cases of use of MALDI-TOF analysis of DNA sequencing can be confirmed with the same results, the above results of this study, we hope to establish by MALDI-TOF common in Taiwan Analysis of the Hemoglobin variation database for clinical screening as a future alternative variant hemoglobin. |
