Evaluation of microchip electrophoresis as a molecular diagnostic method for Duchenne muscular dystrophy.
Evaluation of microchip electrophoresis as a molecular diagnostic method for Duchenne muscular dystrophy.
复制标题
微芯片电泳作为杜氏肌营养不良症分子诊断方法的评价。
DOI：
10.1093/clinchem/48.2.380
复制
发表时间：
2002
影响因子：
9.3
通讯作者：
J. Landers
中科院分区：
文献类型：
--
作者：
J. Ferrance;K. Snow;J. Landers
杜氏肌营养不良症（DMD）是一种由X染色体突变引起的遗传性疾病，目前通过Southern印迹分析进行诊断。受影响的男性和女性携带者通过检测dystropin基因中重复或缺失的外显子来识别。该方法的缺点包括杂交所需的大量片段以确定基因组改变以及分析的耗时、昂贵和繁琐的性质。然而，在DMD中观察到的缺失/重复倾向于位于基因的某些"热点"区域，允许通过询问有限数量的PCR扩增的DNA片段来容易地检测大量病例。
Chamberlain及其同事（1）（2）开发了引物，用于在单一多重PCR扩增中从已知缺失位点扩增9个片段。Beggs等人（3）定义了在第二次多重PCR中扩增9个片段的额外引物。这总共18个片段的集合允许通过PCR快速且准确地检测通过Southern印迹分析可检测的> 97%的缺失。
PCR片段的分析最常在平板凝胶上进行，但正确大小的片段的存在和量都很重要，因为载体具有突变和野生型基因。许多提供DMD检测的临床实验室不提供携带者检测，因为与定量分析相关的技术困难。为了加速分析和自动定量，几个研究小组已经利用毛细管电泳（CE）分离DMD相关片段（4）（5）（6）。其中一个报告通过CE检测DMD的小组将他们的方法转移到电泳微设备（6），这是一种非常快速的分离技术，被证明是用于临床分析的有效工具（7）（8）（9）（10）（11）（12）（13）。尽管这些报告显示CE和微芯片电泳...
Duchenne muscular dystrophy (DMD), a genetic disease caused by mutations in the X chromosome, is currently diagnosed by Southern blot analysis. Affected males and female carriers are identified by the detection of duplicated or deleted exons in the dystropin gene. Disadvantages of this method include the large number of fragments required for hybridization to determine genomic alterations and the time-consuming, expensive, and tedious nature of the analysis. The deletions/duplications seen in DMD tend to be located at certain “hot spot” regions of the gene, however, allowing easy detection of a great number of cases by interrogating a limited number of PCR-amplified DNA fragments.
Chamberlain and coworkers (1)(2) developed primers to amplify nine fragments from known deletion sites in a single multiplex PCR amplification. Beggs et al. (3) defined additional primers that amplify nine fragments in a second multiplex PCR. This total set of 18 fragments allows >97% of the deletions detectable by Southern blot analysis to be detected rapidly and accurately by PCR.
Analysis of the PCR fragments is most frequently carried out on slab gels, but both the presence and amount of the correctly sized fragments are important because carriers have both the mutated and wild-type gene. Many clinical laboratories that offer DMD testing do not provide carrier testing because of the technical difficulties associated with quantitative analysis. To speed analysis and automate quantification, several groups have exploited capillary electrophoresis (CE) for the separation of DMD-relevant fragments (4)(5)(6). One of the groups that reported DMD detection by CE transferred their method to an electrophoretic microdevice (6), a very rapid separation technique demonstrated to be an effective tool for use in clinical analyses (7)(8)(9)(10)(11)(12)(13). Although these reports show CE and microchip electrophoresis …
影响因子：
4.4
作者：
Tian, HJ;Jaquins-Gerstl, A;Landers, JP
通讯作者：
Landers, JP
DOI：
10.1042/bj3280159
发表时间：
1997-11
期刊：
The Biochemical journal
影响因子：
--
作者：
Chad D. Listrom;H. Morizono;Basavapatna S. Rajagopal;Mark T. McCANN;Mendel Tuchman;N. Allewell
通讯作者：
Chad D. Listrom;H. Morizono;Basavapatna S. Rajagopal;Mark T. McCANN;Mendel Tuchman;N. Allewell
微型电泳装置的分子诊断：从平板凝胶到毛细管再到基于微芯片的 T 和 B 细胞淋巴增殖性疾病检测。
影响因子：
9.3
作者：
Munro,NJ;Snow,K;Kant,JA;Landers,JP
通讯作者：
Landers,JP