Extensive screening system using suspension array technology to detect mitochondrial DNA point mutations.
Extensive screening system using suspension array technology to detect mitochondrial DNA point mutations.
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DOI：
10.1016/j.mito.2010.01.003
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发表时间：
2010-04
期刊：
影响因子：
4.4
通讯作者：
Tanaka M
中科院分区：
文献类型：
--
作者：
Nishigaki Y;Ueno H;Coku J;Koga Y;Fujii T;Sahashi K;Nakano K;Yoneda M;Nonaka M;Tang L;Liou CW;Paquis-Flucklinger V;Harigaya Y;Ibi T;Goto Y;Hosoya H;DiMauro S;Hirano M;Tanaka M
我们建立了一个广泛而快速的系统，使用悬浮阵列检测22个基因的61个代表性线粒体DNA （mtDNA）异质或同质点突变（A系列29个，B系列32个）：MT-RNR1、-TV、-ND1、-TQ、-TW、-TC和-TH基因各1个；MT-TN、-TG、-ND4、-TL2、-TE、-CYB基因各2个；MT-ATP6、-ND3和-ND5基因各3个；MT-CO1和-TK基因各4个；MT-TI、-TS1和-ND6基因各5个；MT-TL1基因中有10个。我们仔细选择了5 ' -生物素化引物，并汇集了引物用于两组多重pcr扩增。为了检测突变型和野生型mtDNA，即使在目标突变位点附近存在多态性，我们设计了特定的寡核苷酸探针。利用A系列（29个突变）和B系列（32个突变）的mtDNA点突变检测系统，在107份A系列DNA样本中筛选出3103个突变位点，在397份B系列DNA样本中筛选出13101个突变位点，成功确定了99.4% （A系列）和99.6% （B系列）的靶向突变位点。在该检测系统中，22份携带m.3243A>G异质突变的样品，突变型和野生型特异性探针均显示阳性信号，检测限约为2%。这个基因筛选平台是有用的，以达到明确的诊断线粒体疾病。
We established an extensive and rapid system using suspension array to detect 61 representative mitochondrial DNA (mtDNA) heteroplasmic or homoplasmic point mutations (29 for Series A and 32 for Series B) in 22 genes: 1 each in MT-RNR1, -TV, -ND1, -TQ, -TW, -TC, and -TH genes; 2 each in MT-TN, -TG, -ND4, -TL2, -TE, and -CYB genes; 3 each in MT-ATP6, -ND3, and -ND5 genes; 4 each in MT-CO1 and -TK genes; 5 each in MT-TI, -TS1, and -ND6 genes; and 10 in the MT-TL1 gene. We carefully selected 5′-biotinylated primers and pooled primers for use in two sets of multiplex-PCR amplifications. To detect both mutant and wild-type mtDNA, even when polymorphisms were present near the target mutation sites, we designed specific oligonucleotide probes. By using the mtDNA point mutation detection system of Series A (29 mutations) and Series B (32 mutations), we screened a total of 3103 mutant sites in 107 DNA samples for Series A and 13,101 mutant sites in 397 DNA samples for Series B. We succeeded in determining 99.4% (Series A) and 99.6% (Series B) of the targeted mutant sites by use of the system. The 22 samples with the m.3243A>G heteroplasmic mutation revealed positive signals with both mutant- and wild-type-specific probes in this detection system with a detection limit of approximately 2%. This genetic screening platform is useful to reach a definitive diagnosis for mitochondrial diseases.
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影响因子：
30.8
作者：
PREZANT, TR;AGAPIAN, JV;FISCHELGHODSIAN, N
通讯作者：
FISCHELGHODSIAN, N
影响因子：
64.8
作者：
GOTO, Y;NONAKA, I;HORAI, S
通讯作者：
HORAI, S
影响因子：
5.2
作者：
Leveque, Marianne;Marlin, Sandrine;Denoyelle, Francoise
通讯作者：
Denoyelle, Francoise
影响因子：
4.4
作者：
Bannwarth, S.;Procaccio, V.;Paquis-Flucklinger, V.
通讯作者：
Paquis-Flucklinger, V.
影响因子：
9.8
作者：
Elliott, Hannah R.;Samuels, David C.;Eden, James A.;Relton, Caroline L.;Chinnery, Patrick F.
通讯作者：
Chinnery, Patrick F.