T790M Mutation Test Kit

It has recently been reported that in a proportion of recurrent tumours the presence of a secondary mutation in the EGFR kinase domain, T790M, confers drug resistance. The cytosine to thymidine change is predicted to change threonine to methionine in the catalytic site of the tyrosine kinase domain. These tumours demonstrate sensitivity to a class of irreversible inhibitors of T790M-mutant EGFR.

The T790M mutation kit consists of two real-time PCR assays, one to detect the mutant base at position 2369 of the EGFR gene and a control assay to quantify the EGFR gene. The control assay is designed in a quiet exon (in terms of mutation hotspots). Discrimination is by ARMS® primers and Scorpions® real-time PCR technology is responsible for the  fluorescent detection of PCR products.

 

Intended Use

The kit is sold as a research tool to enable researchers to understand the association between T790M mutations and progression of lung cancers. It will also be useful for drug companies who wish to know the T790M status of patients in clinical trials.

Technological Principles

The T790M test uses a combination of Scorpions® and ARMS® (allele specific PCR) technology. This approach allows the development of very sensitive tests that can detect mutations in a background of normal cells. The real-time PCR based test enables rapid identification and quantification of the mutations. Mutations can be detected at a ratio of 1:100 mutant: normal DNA and this allows the kit to detect genetic variation that could not be detected using DNA sequencing methods.

Protocol

After DNA extraction, real time PCR assays are performed to detect the target molecule.  By comparing control and mutant sample reactions users can detect and estimate low levels of mutation. No further sample processing is necessary and the time to result is <3 hours.

Key Features

  • Detects T790M mutation in exon 20 of the EGFR gene.
  • Qualitative and Quantitative: the proportion of mutant may be measured by comparison to the supplied control.
  • Optimised to detect mutated sequences in an excess of normal DNA. Lower limit of detection is 0.1%.
  • Based on validated technologies; Scorpions for real-time detection and ARMS (allele specific PCR) for selective and specific amplification of mutations.
  • Compatible with most real-time PCR instruments.
  • Sample types include DNA from frozen or paraffin embedded tissue.
  • This kit is for research use only.

 
Kit is available in two different kit sizes:

25 reaction kit              Product Code: TM-01
100 reaction kit            Product Code: TM-02

To place an order please email sales@dxsgenotyping.com or call +44 161 606 7201 for further information or prices.

 
References

  1. Lynch TJ, Bell DW, Sordella R, et al. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med 2004;350:2129-39.

  2. Paez JG, Janne PA, Lee JC, et al. EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science 2004;304:1497-500.

  3. Kobayashi, S., Boggon, T. J., Dayaram, T., et al. EGFR Mutation and Resistance of Non-Small-Cell Lung Cancer to Gefitinib. N Engl J Med 2005;352:786-792.

  4. Kwak EL, Sordella R, Bell DW, et al. Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to gefitinib. PNAS 2005;vol.102:no. 21:7665-70

  5. Pao W., Miller V.A., Politi K.A., et al. Acquired Resistance of Lung Adenocarcinoma to Gefitinib or Erlotinib is Associated with a Second Mutation in the EGFR Kinase Domain. 2005. PloS Medicine 2(3) p1-11.

 


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