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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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A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories. / Akimoto, Chizuru; Volk, Alexander; van Blitterswijk, Marka; Van den Broeck, Marleen; Leblond, Claire S; Lumbroso, Serge; Camu, William; Neitzel, Birgit; Onodera, Osamu; van Rheenen, Wouter; Pinto, Susana; Weber, Markus; Smith, Bradley; Proven, Melanie; Talbot, Kevin; Keagle, Pamela; Chesi, Alessandra; Ratti, Antonia; van der Zee, Julie; Alstermark, Helena; Birve, Anna; Calini, Daniela; Nordin, Angelica; Tradowsky, Daniela C; Just, Walter; Daoud, Hussein; Angerbauer, Sabrina; DeJesus-Hernandez, Mariely; Konno, Takuya; Lloyd-Jani, Anjali; de Carvalho, Mamede; Mouzat, Kevin; Landers, John E; Veldink, Jan H; Silani, Vincenzo; Gitler, Aaron D; Shaw, Christopher E; Rouleau, Guy A; van den Berg, Leonard H; Van Broeckhoven, Christine; Rademakers, Rosa; Andersen, Peter M; Kubisch, Christian.

In: J MED GENET, Vol. 51, No. 6, 01.06.2014, p. 419-24.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Akimoto, C, Volk, A, 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 2014, 'A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories', J MED GENET, vol. 51, no. 6, pp. 419-24. https://doi.org/10.1136/jmedgenet-2014-102360

APA

Akimoto, C., Volk, A., van Blitterswijk, M., Van den Broeck, M., Leblond, C. S., 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., ... Kubisch, C. (2014). A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories. J MED GENET, 51(6), 419-24. https://doi.org/10.1136/jmedgenet-2014-102360

Vancouver

Akimoto C, Volk A, van Blitterswijk M, Van den Broeck M, Leblond CS, Lumbroso S et al. A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories. J MED GENET. 2014 Jun 1;51(6):419-24. https://doi.org/10.1136/jmedgenet-2014-102360

Bibtex

@article{4d1d0e108db441849ab1c17df2980e24,
title = "A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories",
abstract = "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.",
author = "Chizuru Akimoto and Alexander Volk and {van Blitterswijk}, Marka and {Van den Broeck}, Marleen and Leblond, {Claire S} and Serge Lumbroso and William Camu and Birgit Neitzel and Osamu Onodera and {van Rheenen}, Wouter and Susana Pinto and Markus Weber and Bradley Smith and Melanie Proven and Kevin Talbot and Pamela Keagle and Alessandra Chesi and Antonia Ratti and {van der Zee}, Julie and Helena Alstermark and Anna Birve and Daniela Calini and Angelica Nordin and Tradowsky, {Daniela C} and Walter Just and Hussein Daoud and Sabrina Angerbauer and Mariely DeJesus-Hernandez and Takuya Konno and Anjali Lloyd-Jani and {de Carvalho}, Mamede and Kevin Mouzat and Landers, {John E} and Veldink, {Jan H} and Vincenzo Silani and Gitler, {Aaron D} and Shaw, {Christopher E} and Rouleau, {Guy A} and {van den Berg}, {Leonard H} and {Van Broeckhoven}, Christine and Rosa Rademakers and Andersen, {Peter M} and Christian Kubisch",
note = "Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.",
year = "2014",
month = jun,
day = "1",
doi = "10.1136/jmedgenet-2014-102360",
language = "English",
volume = "51",
pages = "419--24",
journal = "J MED GENET",
issn = "0022-2593",
publisher = "BMJ PUBLISHING GROUP",
number = "6",

}

RIS

TY - JOUR

T1 - A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories

AU - Akimoto, Chizuru

AU - Volk, Alexander

AU - van Blitterswijk, Marka

AU - Van den Broeck, Marleen

AU - Leblond, Claire S

AU - Lumbroso, Serge

AU - Camu, William

AU - Neitzel, Birgit

AU - Onodera, Osamu

AU - van Rheenen, Wouter

AU - Pinto, Susana

AU - Weber, Markus

AU - Smith, Bradley

AU - Proven, Melanie

AU - Talbot, Kevin

AU - Keagle, Pamela

AU - Chesi, Alessandra

AU - Ratti, Antonia

AU - van der Zee, Julie

AU - Alstermark, Helena

AU - Birve, Anna

AU - Calini, Daniela

AU - Nordin, Angelica

AU - Tradowsky, Daniela C

AU - Just, Walter

AU - Daoud, Hussein

AU - Angerbauer, Sabrina

AU - DeJesus-Hernandez, Mariely

AU - Konno, Takuya

AU - Lloyd-Jani, Anjali

AU - de Carvalho, Mamede

AU - Mouzat, Kevin

AU - Landers, John E

AU - Veldink, Jan H

AU - Silani, Vincenzo

AU - Gitler, Aaron D

AU - Shaw, Christopher E

AU - Rouleau, Guy A

AU - van den Berg, Leonard H

AU - Van Broeckhoven, Christine

AU - Rademakers, Rosa

AU - Andersen, Peter M

AU - Kubisch, Christian

N1 - Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

PY - 2014/6/1

Y1 - 2014/6/1

N2 - 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.

AB - 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.

U2 - 10.1136/jmedgenet-2014-102360

DO - 10.1136/jmedgenet-2014-102360

M3 - SCORING: Journal article

C2 - 24706941

VL - 51

SP - 419

EP - 424

JO - J MED GENET

JF - J MED GENET

SN - 0022-2593

IS - 6

ER -