A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories

被引：108

作者：

Akimoto, Chizuru ^{[1
]}

Volk, Alexander E. ^{[2
]}

van Blitterswijk, Marka ^{[3
]}

Van den Broeck, Marleen ^{[4
,5
]}

Leblond, Claire S. ^{[6
]}

Lumbroso, Serge ^{[7
]}

Camu, William ^{[8
]}

Neitzel, Birgit ^{[9
]}

Onodera, Osamu ^{[10
]}

van Rheenen, Wouter ^{[11
]}

Pinto, Susana ^{[12
]}

Weber, Markus ^{[13
,14
]}

Smith, Bradley ^{[15
,16
]}

Proven, Melanie ^{[17
]}

Talbot, Kevin ^{[18
]}

Keagle, Pamela ^{[19
]}

Chesi, Alessandra ^{[20
]}

Ratti, Antonia ^{[21
,22
,23
]}

van der Zee, Julie ^{[5
]}

Alstermark, Helena ^{[1
]}

Birve, Anna ^{[1
]}

Calini, Daniela ^{[21
,22
,23
]}

Nordin, Angelica ^{[1
]}

Tradowsky, Daniela C. ^{[2
]}

Just, Walter ^{[2
]}

Daoud, Hussein ^{[6
]}

Angerbauer, Sabrina ^{[9
]}

DeJesus-Hernandez, Mariely ^{[3
]}

Konno, Takuya ^{[10
]}

Lloyd-Jani, Anjali ^{[17
]}

de Carvalho, Mamede ^{[12
]}

Mouzat, Kevin

Landers, John E. ^{[19
]}

Veldink, Jan H. ^{[11
]}

Silani, Vincenzo ^{[21
,22
,23
]}

Gitler, Aaron D. ^{[20
]}

Shaw, Christopher E. ^{[15
,16
]}

Rouleau, Guy A. ^{[6
]}

van den Berg, Leonard H. ^{[11
]}

Van Broeckhoven, Christine ^{[4
,5
]}

Rademakers, Rosa

Andersen, Peter M. ^{[1
,24
]}

Kubisch, Christian ^{[2
]}

机构：

[1] Umea Univ, Dept Pharmacol & Clin Neurosci, SE-90185 Umea, Sweden

[2] Univ Ulm, Inst Human Genet, D-89069 Ulm, Germany

[3] Mayo Clin, Dept Neurosci, Jacksonville, FL 32224 USA

[4] Univ Antwerp VIB, CDE, Dept Mol Genet, Neurodegenerat Brain Dis Grp, B-2610 Antwerp, Belgium

[5] Univ Antwerp, Inst Born Bunge, Neurogenet Lab, Diagnost Serv Facil, B-2020 Antwerp, Belgium

[6] McGill Univ, Montreal Neurol Inst & Hosp, Dept Neurol & Neurosurg, Montreal, PQ, Canada

[7] Univ Nimes Hosp, Dept Biochem, Nimes 9, France

[8] Hop Gui de Chauliac, Montpellier Univ Hosp, Ctr SLA, Montpellier 5, France

[9] Med Genet Zentrum, Munich, Germany

[10] Niigata Univ, Brain Res Inst, Dept Neurol, Niigata 951, Japan

[11] Univ Med Ctr Utrecht, Brain Ctr Rudolf Magnus, Dept Neurol, Utrecht, Netherlands

[12] Univ Lisbon, Fac Med, Inst Mol Med, Hosp Santa Maria, P-1699 Lisbon, Portugal

[13] Kantonsspital St Gallen, Dept Neurol, St Gallen, Switzerland

[14] Univ Hosp, St Gallen, Switzerland

[15] Kings Coll London, Inst Psychiat, London, England

[16] Kings Hlth Partners, London, England

[17] Churchill Hosp, Oxford Med Genet Labs, Oxford OX3 7LJ, England

[18] Univ Oxford, John Radcliffe Hosp, Nuffield Dept Clin Neurosci, Oxford OX3 9DU, England

[19] Univ Massachusetts, Sch Med, Dept Neurol, Worcester, MA USA

[20] Stanford Univ, Dept Genet, Sch Med, Stanford, CA 94305 USA

[21] Univ Milan, Dino Ferrari Ctr, Dept Pathophysiol & Transplantat, Milan, Italy

[22] IRCCS Ist Auxol Italiano, Dept Neurol, Milan, Italy

[23] IRCCS Ist Auxol Italiano, Lab Neurosci, Milan, Italy

[24] Univ Ulm, Dept Neurol, D-89069 Ulm, Germany

来源：

JOURNAL OF MEDICAL GENETICS | 2014年 / 51卷 / 06期

关键词：

FRAGILE-X-SYNDROME; HEXANUCLEOTIDE REPEAT; IDENTIFICATION; CARRIERS; COHORT; FTD; ALS;

D O I：

10.1136/jmedgenet-2014-102360

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Background 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. Methods 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. Results 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. Conclusions 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.

引用

页码：419 / 424

页数：6

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