Detection of DNA mutations associated with mitochondrial diseases by Agilent 2100 bioanalyzer
Detection of DNA mutations associated with mitochondrial diseases by Agilent 2100 bioanalyzer
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DOI：
10.1016/s0009-8981(01)00809-9
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发表时间：
2002-04-01
影响因子：
5
通讯作者：
Wei, YH
中科院分区：
文献类型：
--
作者：
Lu, CY;Tso, DJ;Wei, YH
背景：线粒体DNA （mtDNA）的分子分析为许多线粒体疾病提供了最终诊断。我们对Agilent 2100生物分析仪（Agilent Technologies, Palo Alto， CA）进行了评估，以确定该系统是否可以取代琼脂糖凝胶电泳的传统限制性片段长度多态性（RFLP）分析，用于检测mtDNA突变。方法：本研究招募了3名MELAS综合征家庭成员和4名MERRF综合征家庭成员。PCR和限制性内切酶切后，DNA片段在Agilent 2100生物分析仪上与DNA 500和DNA 1000 Labchip试剂盒一起分离，并在预制的3%琼脂糖凝胶上电泳。结果：与传统方法相比，使用Agilent 2100生物分析仪进行DNA 500和DNA 1000测定的数据具有更低的误差百分比和更好的重复性。结论：基于生物分析仪的性能，我们认为这种新型Labchip足以取代目前琼脂糖凝胶电泳的RFLP分析用于mtDNA突变检测。(C) 2002 Elsevier Science B.V.版权所有
Background: Molecular analysis of mitochondrial DNA (mtDNA) has provided a final diagnosis for many of the mitochondrial diseases. We evaluated the Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA) to determine whether the system could replace the conventional restriction fragment length polymorphism (RFLP) analysis by the agarose gel electrophoresis for the detection of the mtDNA mutation. Methods: Three members of a family with MELAS syndrome and four members of a family with MERRF syndrome were recruited for this study. After PCR and restriction enzyme digestion, DNA fragments were separated on the Agilent 2100 bioanalyzer in conjunction with the DNA 500 and DNA 1000 Labchip kits and by electrophoresis on precast 3% agarose gels. Results: The data generated by the DNA 500 and DNA 1000 assays using the Agilent 2100 bioanalyzer showed a lower percentage error and a better reproducibility as compared to those obtained by the conventional method. Conclusion: Based on the performance of the bioanalyzer, we suggest that this novel Labchip is adequate to replace the current RFLP analysis by the agarose gel electrophoresis for mtDNA mutation detection. (C) 2002 Elsevier Science B.V. All rights reserved.