Signature KRAS Mutations 7 (RUO)* - Theradiag · 877.777.1874 • 2500-0220REV2 Excellent...
Transcript of Signature KRAS Mutations 7 (RUO)* - Theradiag · 877.777.1874 • 2500-0220REV2 Excellent...
877.777.1874 • www.asuragen.com
*�Research�Use�Only.�Not�for�use�in�diagnostic�procedures.
Multiplex assay for 7 distinct mutations in KRAS codon 12 and 13
Co-detection of a KRAS endogenous control sequence in the same reaction
Positive and Negative control samples provided independently
No cross reactivity between mutations and automatic detection on the Luminex® 100 ISTM or 200TM systems
Simultaneous detection and identification in a single reaction well of a 96-well plate
Assess DNA quality and integrity in each DNA specimen
Assess validity of the PCR, hybridization and detection steps in each run
Easy to interpret qualitative results
KRAS�is�an�oncogene�involved�in�the�epidermal�growth�factor�receptor�(EGFR)�signaling�pathway�that�controls�cancer�cell�proliferation�and�apoptosis.�Recently,�there�has�been�increased�interest�in�the�use�of�specific�biomarkers�in�colorectal�cancer�(CRC)�and�other�cancers�treated�with�EGFR�targeted�therapies.�The�Signature®�KRAS�Mutations�7�(RUO)*�assay�is�based�on�the�Signature®�Technology�Platform�for�the�rapid�multiplex�analysis�of�nucleic�acid�sequences,�allowing�detection�of�up�to�100�DNA�or�RNA�targets�in�a�single�reaction�well.�The�Signature®�KRAS�Mutations�(RUO)*�provides�comprehensive�information�on�7�distinct�mutations�within�codons�12�and�13�of�the�KRAS�oncogene.�The�96-well�assay�format�enables�characterization�of�KRAS�mutation�status�in�approximately�4�hours�using�a�broad�input�range�of�genomic�DNA�isolated�from�cultured�cells�or�fresh,�frozen�or�formalin-fixed�paraffin-embedded�(FFPE)�specimens.�The�streamlined�workflow�and�multiplex�assay�format�increase�specimen�throughput,�decrease�reagents/supplies�use�and�reaction�set�up�time,�and�overall,�reduce�the�cost�per�individual�mutation�tested.
The�same�technology�is�also�available�for�detection�of�12�KRAS�mutations�and�BRAF�V600E�as�part�of�the�Signature®�KRAS/BRAF�Mutations�(RUO)*�Assay.
Signature® KRAS Mutations 7 (RUO)*
Figure 1:�Seven�distinct�KRAS�mutations�can�be�detected�by�the�assay�in�a�single�reaction�(right).�The�streamlined�assay�workflow�is�optimized�for�rapid�time�to�results�with�minimum�hands-on�time�(above).
G12S� (GGT>AGT)G12R� (GGT>CGT)G12C� (GGT>TGT)G12D� (GGT>GAT)G12A� (GGT>GCTG12V� (GGT>GTT)G13D� (GGC>GAC)
Key Attributes of Signature® KRAS Mutations 7 (RUO)*:
877.777.1874 • www.asuragen.com
2500-0220REV2
Excellent analytical specificity across all 7 mutations using the Signature® KRAS Mutations 7 Assay*
At least 1% analytical sensitivity using the Signature® KRAS Mutations 7 Assay*
Kit Negative control
Kit Positive control
No DNA Control
Cell line WT (HT-29)
Cell line G12V (HOM)
Cell line G12A (HET)
Cell line G12D (HET)
Cell line G12C (HOM)
Cell line G12S (HET)
Cell line G13D (HET)
Plasmid G12R in HT-29
G12V, GTT
117
3771
142
153
5202
135
169
186
167
240
197
G12A, GCT
106
206
258
171
315
6732
173
186
164
147
245
G12D, GAT
96
105
127
133
106
367
5317
84
125
158
184
G12C, TGT
105
15
108
126
114
128
87
5180
141
93
221
G12S, AGT
169
169
165
213
211
135
94
498
3845
189
181
G13D, GAC
162
172
139
81
83
125
135
53
112
4245
411
G12R, CGT
176
207
120
218
222
102
191
214
87
112
3537
KEC
4912
5292
118
8608
8994
8803
8656
8483
9125
8497
7986
Sample
Signature® KRAS Mutations 7 (RUO)*
Figure 2:�Representative�assay�MFI�output�data�with�10�ng�of�genomic�DNA�purified�from�7�cultured�cell�lines�either�wild�type�(WT)�for�KRAS�or�carrying�specific�homozygous�(HOM)�or�heterozygous�(HET)�KRAS�mutations.�Detection�of�G12R�mutation�target�was�tested�using�a�plasmid�DNA�carrying�the�GGT>CGT�mutation�(5,000�copies�spiked�into�10�ng�of�WT-HT-29�cell�line�DNA).�KEC�=�KRAS�Endogenous�Control.�
Figure 3:�Representative�examples�of�analytical�sensitivity�with�genomic�DNA�purified�from�a�homozygous�G12V�positive�cell�line�or�a�heterozygous�G13D�positive�cell�line.�The�DNA�samples�were�either�undiluted�(100%)�or�diluted�in�a�background�of�wild�type�genomic�DNA�(HT-29)�keeping�the�total�input�constant�at�20�ng.�A�sensitivity�equivalent�to�at�least�1%�(1�ng�of�positive�DNA�in�19�ng�of�WT�DNA)�was�achieved.
References:1�Walther�A,�Johnstone�E,�Swanton�C,�Midgley�R,�Tomlinson�I,�and�Kerr�D.�Genetic�prognostic�and�predictive�markers�in�colorectal�cancer.�Nat�Rev�Cancer.�2009�Jul;9(7):489-99.2�Linardou�H,�Dahabreh�IJ,�Bafaloukos�D,�Kosmidis�P,�and�Murray�S.�Somatic�EGFR�mutations�and�efficacy�of�tyrosine�kinase�inhibitors�in�NSCLC.�Nat�Rev�Clin�Oncol.�2009�Jun;6(6):352-66.
[P/N 49418] 48 Reactions[P/N 49419] 48 Reactions
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*�Preliminary�research�data,�the�performance�characteristics�of�this�assay�are�not�yet�established.
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