Denaturing High Performance Liquid Chromatography Screening of Ryanodine Receptor Type 1 Gene in Patients with Malignant Hyperthermia in Taiwan and Identification of a Novel Mutation (Y522C)
Denaturing High Performance Liquid Chromatography Screening of Ryanodine Receptor Type 1 Gene in Patients with Malignant Hyperthermia in Taiwan and Identification of a Novel Mutation (Y522C)
复制标题
台湾恶性高热患者瑞尼定受体1型基因变性高效液相色谱筛查及新突变（Y522C）的鉴定
作者：
H. Yeh;Mei;Yidi Su;Ronghuan Shen;J. Hwang;Wei;Ling
本研究旨在鉴定台湾恶性高热(MH)患者的基因突变。我们还测试了变性高效液相色谱(DHPLC)方案可用于这些患者的突变检测的假设。我们发现五位台湾患者在全身麻醉后有典型的MH临床表现。我们还招募了50名健康志愿者。聚合酶链式反应扩增兰尼定受体(RYR1)基因突变热点，DHPLC技术筛查突变。在DHPLC分析中检测到杂合子洗脱模式后，进行DNA测序反应以鉴定核苷酸变异。我们在所有5例MH患者中都发现了RYR1突变。在5例患者中发现4种不同的突变：Tyr522Cys、Arg552Trp、Val2168Met和Thr2206Arg。在5例患者中，2例无关患者具有相同的Thr2206Arg突变。其中三个突变以前已经报道过，但Tyr522Cys突变是新的。对照组未发现MH相关突变。综上所述，我们使用基于DHPLC的方法在5名台湾MH患者中发现了RYR1突变。基于DHPLC的基因检测有可能成为MH的一种无创、简便的检测方法。
We performed the present study to identify the mutation in patients in Taiwan with malignant hyperthermia (MH). We also test the hypothesis that a denaturing high-performance liquid chromatography (DHPLC) protocol can be used for mutation detection in these patients. We identified five Taiwanese patients with typical clinical presentations of MH after general anesthesia. We also enrolled 50 healthy volunteers. Polymerase chain reaction was used to amplify the ryanodine receptor (RYR1) gene mutation hot spots and DHPLC techniques were used to screen for mutations. Upon detection of a heterozygous elution pattern in DHPLC analysis, DNA sequencing reaction was performed to identify the nucleotide variations. We identified a RYR1 mutation in all 5 patients with MH. There were 4 different mutations in the 5 patients: Tyr522Cys, Arg552Trp, Val2168Met, and Thr2206Arg. Among the 5 patients, 2 unrelated patients had the same Thr2206Arg mutation. Three of the mutations had been reported before, but the Tyr522Cys mutation was novel. None of the MH-related mutations were found in the control group. In conclusion, we identified RYR1 mutations in 5 Taiwanese patients with MH using a DHPLC-based approach. A DHPLC-based genetic test may be developed as a noninvasive and convenient test for MH.