AKT1 E17K Mutation Analysis Kit

Available Products

Product Name/Description
No. of Reactions*
Product Code
CRCdx® RAS Mutation Detection Kit
40
 CRCDX-RAS

For in vitro diagnostic use

Intended Use

The CRCdx® RAS Mutation Detection Kit is a qualitative real-time PCR in vitro diagnostic test intended for the detection of 35 variants of KRAS and NRAS exon 2, 3, 4 somatic mutations in genomic DNA extracted from formalin-fixed, paraffin-embedded (FFPE) colorectal cancer (CRC) tissue samples. The test is intended as a companion diagnostic (CDx) to aid in the identification of colorectal cancer (mCRC) patients who may benefit from treatment with Vectibix (panitumumab) based on a no mutation detected test result in accordance with the approved therapeutic product labeling.

Kit Features

Detects somatic mutations found in exons 2, 3, and 4 of KRAS and NRAS genes. Table 1 lists the mutations detected by this kit.

Table 1. List of mutations in KRAS and NRAS detected by CRCdx® RAS Mutation Detection Kit

Gene

Exon

AA Change

Nucleotide Change

Cosmic ID (COSV)

Reaction Mix

KRAS

2

G12A

c.35G>C

55497479

1

G12D

c.35G>A

55497369

G12R

c.34G>C

55497582

G12V

c.35G>T

55497419

G13D

c.38G>A

55497388

G12C

c.34G>T

55497469

2

G12S

c.34G>A

55497461

3

Q61H

c.183A>T

55499223

3

Q61H

c.183A>C

55498802

Q61L

c.182A>T

55504296

Q61R

c.182A>G

55498739

A59E

c.176C>A

55568979

A59G

c.176C>G

55604554

A59T

c.175G>A

55499283

4

K117N(AAC)

c.351A>C

55545304

4

K117N(AAT)

c.351A>T

55504752

K117R

c.350A>G

55937655

K117E

c.349A>G

55716212

A146T

c.436G>A

55501778

5

A146P

c.436G>C

55541748

A146V

c.437C>T

55498939

NRAS

2

G12D

c.35G>A

54736383

6

G12C

c.34G>T

54736487

G12S

c.34G>A

54736621

G13R

c.37G>C

54736550

G13V

c.38G>T

54736480

4

K117R

c.350A>G

104682079

3

Q61H (CAC)

c.183A>C

54736320

7

Q61H (CAT)

c.183A>T

54736991

Q61L

c.182A>T

54736624

Q61K

c.181C>A

54736310

Q61R

c.182A>G

54736340

3 and 4

A146T

c.436G>A

65731978

8

A59D

c.176C>A

54738004

A59T

c.175G>A

54743124

 

Assay Principle

The CRCdx® RAS Mutation Detection Kit is a polymerase chain reaction (PCR)-based assay that uses allele-specific primers in a multiplex reaction to identify the presence of the mutations in a total of 8 reactions per sample. The test does not distinguish between mutations within each group. The reagents in the kit include eight (8) vials of primer/probe mix designed to detect distinct groups of mutations (see kit content table below), five (5) vials of reaction mix (enzyme), and one (1) vial of positive control mix. The nuclease free water for the No Template Control (NTC) or negative control is required but not provided with the kit. The kit allows for 40 specimens to be tested.

The testing procedure involves three (3) simple steps:

  • Isolation of DNA from paraffin-embedded sections (FFPE) using Promega Maxwell® CSC
  • Real-time PCR using ThermoFisher Scientific QuantStudio® Dx real-time PCR instrument
  • Data analysis and interpretation using EntroGen PCR Analysis Software (EPAS™) verison 2.0

This test can be completed in approximately 2 hours from isolation of DNA to test result.

 

Equipment and Materials

  • Promega Maxwell® CSC
  • Promega Maxwell® CSC DNA FFPE Kit
  • ThermoFisher Scientific QuantStudio® Dx real-time PCR instrument

Performance

The safety and effectiveness of the CRCdx RAS Mutation Detection Kit was evaluated in an external concordance study designed to demonstrate non-inferiority to the FDA approved companion diagnostic Praxis® Extended RAS Panel for the purpose of validating the use of the follow-on companion diagnostic test for panitumumab (Vectibix®). The objective of the study was to demonstrate the non-inferiority-equivalence of CRCdx® to Praxis® Extended RAS to infer treatment outcome efficacy of Vectibix® as CRC patients without KRAS or NRAS mutations may benefit from treatment. Archived clinical CRC patient FFPE tissue acquired from biorepositories were tested with CRCdx® RAS and FDA approved Praxis® Extended RAS Panel. A total of 398 were tested. Concordance study was performed to estimate percent agreement between CRCdx® RAS and the Praxis®. Mutation level comparison was performed to assess percent concordance. Concordance analysis demonstrated that CRCdx® has the attributes of PPA = 99.6%, NPA = 92%, and OPA = 96.2%.

 

Table 2. Summary of Positive, Negative, and Overall Percent Agreement for CRCdx® RAS with Praxis

Praxis

PPA

NPA

OPA

Mutant

WT

Total

CRCdx

Mutant

223

14

237

99.6%

92.0%

96.2%

WT

1

160

161

Total

224

174

398

95% CI

97.5% to 100.0%

86.8% to 95.5%

93.9% to 97.9%

Resources

CRCdx® RAS Mutation Detection Kit Instructions for Use

 

FDA.gov – Recently-Approved Devices

 

Information about EPAS™

The following antivirus applications are compatible with EPAS™ version 2.0 Software:

  • Mcafee antivirus versions:
    • 16.0.R52
  • Norton antivirus versions:
    • 22.23.6.5
  • Microsoft Defender Antivirus Security Intelligence versions:
    • 1.397.492.0

Last update: 2023-09-12

Overview

  • FDA-approved for IVD use
  • Works with FFPE
  • Ready to use reagents
  • Includes software for analysis
  • 8 Reactions per sample

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