高肌酸激酶血症新生兒篩檢技術之開發

Development of a newborn screening technique for hyper-creatine kinase-emia

劉麗蘭(Li-Lan Liu)

指導教授 ： 胡務亮

國立臺灣大學/醫學院/分子醫學研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.01935

摘要

背景
肌酸激酶(Creatine kinase, CK)是參與體內能量代謝的一種酵素，也是一種可逆磷酸化的酶，CK可形成高能量的三磷酸腺苷ATP，CK亦能供給肌肉能量的來源。在臨床上，測定血清CK檢查指的是血清中total CK的活性，由於在骨骼肌，心臟肌肉，和腦組織中含有豐富的激酸激酶。因此常用於肌肉相關疾病的檢查，如急性心肌梗塞，當這些部位受傷，血清中的CK會上升。在龐貝氏症、肢帶型肌肉失養症、心肌梗塞患者及肌營養不良症(如裘馨氏肌營養不良症及貝克型肌肉營養不良症)的血清中CK活性均呈現輕度至中度升高。而和CK的相關研究也顯示在裘馨氏肌肉失養症的男性患者CK活性會顯著升高，女性患者的CK活性則會輕微升高。

方法
本研究方法是利用血片檢體，做大規模的篩檢，以免疫螢光法檢測儀檢測血片中肌酸激酶的活性。

結果
本實驗總共分析有8,083個新生兒血片，其中男性有4,178個(51.7 %)，女性有3,905個(48.3%)，男女比例為1.07，在8,083個檢體中，CK值在1500 Flu/min以上共有10個檢體，佔0.12%，其中男性檢體有5人，女性檢體有5人，CK值在1,200至1,500 Flu/min有12個檢體，佔0.15%，男性檢體有7人，女性檢體有5人，CK值在1,000至1,200 Flu/min有23個檢體，佔0.28%，男性檢體有19人，女性檢體有4人。

結論
CK是一種來自肌肉的酵素，因此在肌肉受傷時，會引起CK的上升，本實驗方法是利用新生兒篩檢血片進行大量、快速，且有效的CK活性篩檢，找出高CK疑陽性個案。我們的研究結果，CK Flu/min >= 1,000共計有45個檢體為高疑陽性個案。在CK與其他因子的關連性研究的結果顯示，在CK與性別的關連性上，其在統計上沒有顯著差異(P > 0.05)。在CK與出生體重的關連性上，以體重小於或大於等於2,200 g做分組研究，在統計上具有顯著差異(P < 0.05)。在CK與妊娠週數的關連性上，以妊娠週數小於或大於等於37週做分組研究，在統計上具有顯著差異(P < 0.05)。在CK與CAH的關連性上，以妊娠週數大於等於37週做分組研究，統計上具有顯著差異(P < 0.05)。CK活性的確有可能因生產過程的擠壓而略為升高，但應該不會影響高疑陽性個案的判斷。在CK與性G6PD的關連性上，結果顯示G6PD缺乏症患者的CK數值並不會有偏低之情形發生。G6PD缺乏症應該不會影響CK反應下降，造成偽陰性的判斷。

關鍵字

肌酸激酶 ； 裘馨氏肌肉失養症 ； 代謝性肌肉疾病 ； 新生兒篩檢 ； 乾燥血片

並列摘要

Background
Creatine kinase, a reversible phosphorylation enzyme, is one of the enzymes involved in energy metabolism. It can form a high energy triphosphate ATP and provide the muscle energy. Clinically, serum CK examination refers to the total CK activity in serum. Due to the significant amount of kinase in skeletal and heart muscle, and brain.CK is commonly used as an indicator to examine the muscle-related disorders. When injuries of muscle, brain and heart occur, the CK level is usually elevated. In most patients with Pompe disease, limb-girdle muscular dystrophy, myocardial infarction, and muscular dystrophy (such as Duchenne muscular dystrophy and Becker muscular dystrophy), CK activity also showed mild to moderate elevations. In the cases of Duchenne muscular dystrophy, the serum CK activity is significantly increased in male patients but only slightly elevated in female patients.

Method
Our research method use the dried blood spot samples for mass screening to detect creatine kinase activity by fluorescence detector .

Results
In this study, a total of 8,083 newborns dried blood spots were analyzed. Among them, there were 4,178 male (51.7%) and 3,905 female (48.3%), and the male to female ratio was 1.07. The CK Flu / min values greater than 1,500 or more was detected in 10 cases (0.12%) of which male vs. females were equal. The CK Flu / min values of 1,200-1,500 was found in 12 samples (0.15%)indicated 7 male and 5 female subjects. The CK Flu / min values in 1,000-1,200 was detected in 23 samples (0.28%) 19 male and 4 female subjects.

Conclusions
Clinically, CK is commonly used as an indicator to examine the muscle-related disorders. In this study, we perform a rapid and effective high-through put screening using the newborn screening dried blood spots. From the results, there are 45 cases with higher CK activity (>=1,000 CK Flu/min) in the DBS. In the study of CK and other parameters, we find no relation between CK and Gender, but there are relationship between CK & Birth weight and CK & Gestational age. From the CK and CAH study, birth Stress may certainly make some raise in the CK level, but it may not influence the judgement of hyper-creatine kinase-emia. Although we use the reagents with G6PD enzyme, the G6PD enzyme deficiency in the DBS may not decrease the CK reaction obviously.

並列關鍵字

Creatine kinase ； Duchenne Muscular Dystrophy ； Metabolic muscular disease ； newborn screen ； Dried blood spot

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