Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer

Kazuko Sakai, Azusa Yoneshige, Akihiko Ito, Yoji Ueda, Satoshi Kondo, Hitoshi Nobumasa, Yoshihiko Fujita, Yosuke Togashi, Masato Terashima, Marco A. De Velasco, Shuta Tomida, Kazuto Nishio

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We compared the performance of the 3D-Gene® mutation assay (3D-Gene® KRAS mutation assay kit) with the Scorpion-ARMS (therascreen® KRAS RGQ PCR Kit) and Luminex (MEBGEN™ KRAS kit) assays for the detection of KRAS mutations in formalin-fixed, paraffin-embedded tissue samples from 150 patients diagnosed with colorectal cancer. DNA was extracted from the paraffin-embedded tissue samples with or without macrodissection under hematoxylin and eosin staining and the KRAS mutation status was independently determined using these assays. Discordant results were re-analyzed by Sanger sequencing. Mutation detection analysis was successfully performed in all 150 specimens using the 3D-Gene® mutation assay without an invalid case. The concordance rate between the 3D-Gene® mutation assay and Scorpion-ARMS or Luminex was 98.7% (148/150). KRAS mutations were detected at a frequency of 35.3% (53/150) in colorectal cancer specimens. Three discrepant cases were found between the three assays. Overall, our results demonstrate a high concordance rate of between the 3D-Gene® mutation assay and the two existing in-vitro diagnostics kits. All three assays proved to be validated methods for detecting clinically significant KRAS mutations in paraffin-embedded tissue samples.

Original languageEnglish
Article number7
Pages (from-to)1-6
Number of pages6
JournalSpringerPlus
Volume4
Issue number1
DOIs
Publication statusPublished - Dec 1 2015
Externally publishedYes

Fingerprint

Paraffin
Formaldehyde
Colorectal Neoplasms
Mutation
Scorpions
Genes
Hematoxylin
Eosine Yellowish-(YS)
Staining and Labeling
Polymerase Chain Reaction
DNA

Keywords

  • 3D-Gene® KRAS mutation assay
  • Anti-EGFR antibody
  • Colorectal cancer
  • Companion diagnosis
  • KRAS mutation

ASJC Scopus subject areas

  • General

Cite this

Sakai, K., Yoneshige, A., Ito, A., Ueda, Y., Kondo, S., Nobumasa, H., ... Nishio, K. (2015). Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer. SpringerPlus, 4(1), 1-6. [7]. https://doi.org/10.1186/2193-1801-4-7

Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer. / Sakai, Kazuko; Yoneshige, Azusa; Ito, Akihiko; Ueda, Yoji; Kondo, Satoshi; Nobumasa, Hitoshi; Fujita, Yoshihiko; Togashi, Yosuke; Terashima, Masato; De Velasco, Marco A.; Tomida, Shuta; Nishio, Kazuto.

In: SpringerPlus, Vol. 4, No. 1, 7, 01.12.2015, p. 1-6.

Research output: Contribution to journalArticle

Sakai, K, Yoneshige, A, Ito, A, Ueda, Y, Kondo, S, Nobumasa, H, Fujita, Y, Togashi, Y, Terashima, M, De Velasco, MA, Tomida, S & Nishio, K 2015, 'Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer', SpringerPlus, vol. 4, no. 1, 7, pp. 1-6. https://doi.org/10.1186/2193-1801-4-7
Sakai, Kazuko ; Yoneshige, Azusa ; Ito, Akihiko ; Ueda, Yoji ; Kondo, Satoshi ; Nobumasa, Hitoshi ; Fujita, Yoshihiko ; Togashi, Yosuke ; Terashima, Masato ; De Velasco, Marco A. ; Tomida, Shuta ; Nishio, Kazuto. / Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer. In: SpringerPlus. 2015 ; Vol. 4, No. 1. pp. 1-6.
@article{ec2c19d24f924c41ab6888be4845b2a2,
title = "Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer",
abstract = "We compared the performance of the 3D-Gene{\circledR} mutation assay (3D-Gene{\circledR} KRAS mutation assay kit) with the Scorpion-ARMS (therascreen{\circledR} KRAS RGQ PCR Kit) and Luminex (MEBGEN™ KRAS kit) assays for the detection of KRAS mutations in formalin-fixed, paraffin-embedded tissue samples from 150 patients diagnosed with colorectal cancer. DNA was extracted from the paraffin-embedded tissue samples with or without macrodissection under hematoxylin and eosin staining and the KRAS mutation status was independently determined using these assays. Discordant results were re-analyzed by Sanger sequencing. Mutation detection analysis was successfully performed in all 150 specimens using the 3D-Gene{\circledR} mutation assay without an invalid case. The concordance rate between the 3D-Gene{\circledR} mutation assay and Scorpion-ARMS or Luminex was 98.7{\%} (148/150). KRAS mutations were detected at a frequency of 35.3{\%} (53/150) in colorectal cancer specimens. Three discrepant cases were found between the three assays. Overall, our results demonstrate a high concordance rate of between the 3D-Gene{\circledR} mutation assay and the two existing in-vitro diagnostics kits. All three assays proved to be validated methods for detecting clinically significant KRAS mutations in paraffin-embedded tissue samples.",
keywords = "3D-Gene{\circledR} KRAS mutation assay, Anti-EGFR antibody, Colorectal cancer, Companion diagnosis, KRAS mutation",
author = "Kazuko Sakai and Azusa Yoneshige and Akihiko Ito and Yoji Ueda and Satoshi Kondo and Hitoshi Nobumasa and Yoshihiko Fujita and Yosuke Togashi and Masato Terashima and {De Velasco}, {Marco A.} and Shuta Tomida and Kazuto Nishio",
year = "2015",
month = "12",
day = "1",
doi = "10.1186/2193-1801-4-7",
language = "English",
volume = "4",
pages = "1--6",
journal = "SpringerPlus",
issn = "2193-1801",
publisher = "Springer Science and Business Media Deutschland GmbH",
number = "1",

}

TY - JOUR

T1 - Performance of a novel KRAS mutation assay for formalin-fixed paraffin embedded tissues of colorectal cancer

AU - Sakai, Kazuko

AU - Yoneshige, Azusa

AU - Ito, Akihiko

AU - Ueda, Yoji

AU - Kondo, Satoshi

AU - Nobumasa, Hitoshi

AU - Fujita, Yoshihiko

AU - Togashi, Yosuke

AU - Terashima, Masato

AU - De Velasco, Marco A.

AU - Tomida, Shuta

AU - Nishio, Kazuto

PY - 2015/12/1

Y1 - 2015/12/1

N2 - We compared the performance of the 3D-Gene® mutation assay (3D-Gene® KRAS mutation assay kit) with the Scorpion-ARMS (therascreen® KRAS RGQ PCR Kit) and Luminex (MEBGEN™ KRAS kit) assays for the detection of KRAS mutations in formalin-fixed, paraffin-embedded tissue samples from 150 patients diagnosed with colorectal cancer. DNA was extracted from the paraffin-embedded tissue samples with or without macrodissection under hematoxylin and eosin staining and the KRAS mutation status was independently determined using these assays. Discordant results were re-analyzed by Sanger sequencing. Mutation detection analysis was successfully performed in all 150 specimens using the 3D-Gene® mutation assay without an invalid case. The concordance rate between the 3D-Gene® mutation assay and Scorpion-ARMS or Luminex was 98.7% (148/150). KRAS mutations were detected at a frequency of 35.3% (53/150) in colorectal cancer specimens. Three discrepant cases were found between the three assays. Overall, our results demonstrate a high concordance rate of between the 3D-Gene® mutation assay and the two existing in-vitro diagnostics kits. All three assays proved to be validated methods for detecting clinically significant KRAS mutations in paraffin-embedded tissue samples.

AB - We compared the performance of the 3D-Gene® mutation assay (3D-Gene® KRAS mutation assay kit) with the Scorpion-ARMS (therascreen® KRAS RGQ PCR Kit) and Luminex (MEBGEN™ KRAS kit) assays for the detection of KRAS mutations in formalin-fixed, paraffin-embedded tissue samples from 150 patients diagnosed with colorectal cancer. DNA was extracted from the paraffin-embedded tissue samples with or without macrodissection under hematoxylin and eosin staining and the KRAS mutation status was independently determined using these assays. Discordant results were re-analyzed by Sanger sequencing. Mutation detection analysis was successfully performed in all 150 specimens using the 3D-Gene® mutation assay without an invalid case. The concordance rate between the 3D-Gene® mutation assay and Scorpion-ARMS or Luminex was 98.7% (148/150). KRAS mutations were detected at a frequency of 35.3% (53/150) in colorectal cancer specimens. Three discrepant cases were found between the three assays. Overall, our results demonstrate a high concordance rate of between the 3D-Gene® mutation assay and the two existing in-vitro diagnostics kits. All three assays proved to be validated methods for detecting clinically significant KRAS mutations in paraffin-embedded tissue samples.

KW - 3D-Gene® KRAS mutation assay

KW - Anti-EGFR antibody

KW - Colorectal cancer

KW - Companion diagnosis

KW - KRAS mutation

UR - http://www.scopus.com/inward/record.url?scp=84943641355&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84943641355&partnerID=8YFLogxK

U2 - 10.1186/2193-1801-4-7

DO - 10.1186/2193-1801-4-7

M3 - Article

VL - 4

SP - 1

EP - 6

JO - SpringerPlus

JF - SpringerPlus

SN - 2193-1801

IS - 1

M1 - 7

ER -