A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories.
A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories.
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DOI：
10.1136/jmedgenet-2014-102360
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发表时间：
2014-06
影响因子：
4
通讯作者：
Kubisch C
中科院分区：
文献类型：
--
作者：
Akimoto C;Volk AE;van Blitterswijk M;Van den Broeck M;Leblond CS;Lumbroso S;Camu W;Neitzel B;Onodera O;van Rheenen W;Pinto S;Weber M;Smith B;Proven M;Talbot K;Keagle P;Chesi A;Ratti A;van der Zee J;Alstermark H;Birve A;Calini D;Nordin A;Tradowsky DC;Just W;Daoud H;Angerbauer S;DeJesus-Hernandez M;Konno T;Lloyd-Jani A;de Carvalho M;Mouzat K;Landers JE;Veldink JH;Silani V;Gitler AD;Shaw CE;Rouleau GA;van den Berg LH;Van Broeckhoven C;Rademakers R;Andersen PM;Kubisch C
C9 orf 72中的GGGGCC重复扩增是肌萎缩侧索硬化（ALS）和额颞叶痴呆（FTD）患者中最常见的突变。大多数关于C9 orf 72的研究依赖于重复引物PCR（RP-PCR）方法来检测扩增。为了研究这种技术的固有局限性，我们比较了14个实验室的方法和结果。14个实验室以盲态方式对78名个体（诊断为ALS或FTD）的DNA进行基因分型。11个实验室使用扩增子长度分析和RP-PCR的组合，而3个实验室单独使用RP-PCR; Southern印迹技术用作参考。使用基于PCR的技术，14个实验室中的5个得到的结果与Southern印迹结果完全一致。78份DNA样本中只有50份在所有14个实验室得到相同的基因型结果。存在高度的假阳性和假阴性结果，14个实验室中有9个实验室至少有一个样本根本无法进行基因分型。扩增子长度分析和RP-PCR联合应用的平均敏感性为95.0%（73.9-100%），平均特异性为98.0%（87.5-100%）。总体而言，只有7个实验室观察到超过95%的灵敏度和特异性。由于基因分型结果的范围很广，我们建议在研究环境中至少使用扩增子长度分析和RP-PCR的组合。我们建议Southern印迹技术应该是金标准，并在临床诊断设置强制性。
The GGGGCC-repeat expansion in C9orf72 is the most frequent mutation found in patients with amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Most of the studies on C9orf72 have relied on repeat-primed PCR (RP-PCR) methods for detection of the expansions. To investigate the inherent limitations of this technique, we compared methods and results of 14 laboratories. The 14 laboratories genotyped DNA from 78 individuals (diagnosed with ALS or FTD) in a blinded fashion. Eleven laboratories used a combination of amplicon-length analysis and RP-PCR, whereas three laboratories used RP-PCR alone; Southern blotting techniques were used as a reference. Using PCR-based techniques, 5 of the 14 laboratories got results in full accordance with the Southern blotting results. Only 50 of the 78 DNA samples got the same genotype result in all 14 laboratories. There was a high degree of false positive and false negative results, and at least one sample could not be genotyped at all in 9 of the 14 laboratories. The mean sensitivity of a combination of amplicon-length analysis and RP-PCR was 95.0% (73.9–100%), and the mean specificity was 98.0% (87.5–100%). Overall, a sensitivity and specificity of more than 95% was observed in only seven laboratories. Because of the wide range seen in genotyping results, we recommend using a combination of amplicon-length analysis and RP-PCR as a minimum in a research setting. We propose that Southern blotting techniques should be the gold standard, and be made obligatory in a clinical diagnostic setting.
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影响因子：
3.7
作者：
Dobson-Stone C;Hallupp M;Loy CT;Thompson EM;Haan E;Sue CM;Panegyres PK;Razquin C;Seijo-Martínez M;Rene R;Gascon J;Campdelacreu J;Schmoll B;Volk AE;Brooks WS;Schofield PR;Pastor P;Kwok JB
通讯作者：
Kwok JB
DOI：
10.1016/s1474-4422(12)70043-1
发表时间：
2012-04
期刊：
The Lancet. Neurology
影响因子：
--
作者：
Majounie E;Renton AE;Mok K;Dopper EG;Waite A;Rollinson S;Chiò A;Restagno G;Nicolaou N;Simon-Sanchez J;van Swieten JC;Abramzon Y;Johnson JO;Sendtner M;Pamphlett R;Orrell RW;Mead S;Sidle KC;Houlden H;Rohrer JD;Morrison KE;Pall H;Talbot K;Ansorge O;Chromosome 9-ALS/FTD Consortium;French research network on FTLD/FTLD/ALS;ITALSGEN Consortium;Hernandez DG;Arepalli S;Sabatelli M;Mora G;Corbo M;Giannini F;Calvo A;Englund E;Borghero G;Floris GL;Remes AM;Laaksovirta H;McCluskey L;Trojanowski JQ;Van Deerlin VM;Schellenberg GD;Nalls MA;Drory VE;Lu CS;Yeh TH;Ishiura H;Takahashi Y;Tsuji S;Le Ber I;Brice A;Drepper C;Williams N;Kirby J;Shaw P;Hardy J;Tienari PJ;Heutink P;Morris HR;Pickering-Brown S;Traynor BJ
通讯作者：
Traynor BJ
影响因子：
48
作者：
van Blitterswijk, Marka;Dejesus-Hernandez, Mariely;Niemantsverdriet, Ellis;Murray, Melissa E.;Heckman, Michael G.;Diehl, Nancy N.;Brown, Patricia H.;Baker, Matthew C.;Finch, NiCole A.;Bauer, Peter O.;Serrano, Geidy;Beach, Thomas G.;Josephs, Keith A.;Knopman, David S.;Petersen, Ronald C.;Boeve, Bradley F.;Graff-Radford, Neill R.;Boylan, Kevin B.;Petrucelli, Leonard;Dickson, Dennis W.;Rademakers, Rosa
通讯作者：
Rademakers, Rosa
影响因子：
9.8
作者：
Beck, Jon;Poulter, Mark;Mead, Simon
通讯作者：
Mead, Simon
影响因子：
16.2
作者：
DeJesus-Hernandez M;Mackenzie IR;Boeve BF;Boxer AL;Baker M;Rutherford NJ;Nicholson AM;Finch NA;Flynn H;Adamson J;Kouri N;Wojtas A;Sengdy P;Hsiung GY;Karydas A;Seeley WW;Josephs KA;Coppola G;Geschwind DH;Wszolek ZK;Feldman H;Knopman DS;Petersen RC;Miller BL;Dickson DW;Boylan KB;Graff-Radford NR;Rademakers R
通讯作者：
Rademakers R