| EGFR mutation detection is performed in two tubes. Tube one detects point mutations and tube two detects insertions and deletions. |
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Assay Method: Mutation enrichment by Shifted Termination Assay (STA)
Data collection by fragment analysis
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| Assay platform: Capillary Electrophoresis Sequencer |
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| Assay Time: 2.5 - 3 hours |
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| Assay Throughput: One 96-well plate is able to test 46 samples (including two controls). A 16-capillary sequencer can test up to 150-200 samples per day. |
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| Mutations Detected: |
- Point mutations: E709A, G719S, G719A, S768I, T790M, L858R and L861Q. Can also detect E709V, E709G, G719C, G719D, S768T, L861R, in addition to less frequent mutations in these codons.
- Deletions between codon 745 and codon 759 (>115 deletions are reported in this area by the cosmic database)
- Insertions between codon 765 and codon 775 (>40 insertions are reported in this area by the cosmic database)
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Comparision of EGFR Mutation Detection Methods
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STA Core Reagents |
Real-Time PCR |
Sequencing |
| Mutations Detected |
>120 |
28 |
All mutations |
| Detect T790M |
Yes |
No |
Yes |
| Test Format |
2 tubes |
8 tubes |
>4 tubes |
Throughput
(one 96-well plate) |
46 samples |
10 samples |
12-24 samples |
| Sensitivity |
1% |
1% |
15-20% |
| Data Analysis |
Clear-cut results |
Fluctuating Ct value |
Time consuming analysis |
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| Required Reagents: |
EGFR mutation detection is performed with STA Core Reagents and gene specific primer sets.
Reagent list: |
- STA Core Reagents A Cat. COR-A
- STA Core Reagents C Cat. COR-C
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