Multiplexed diagnostic assay for detection of targetable mutations in lung adenocarcinoma using SNaPshot PCR technique

9^th Indo Global Summit on Cancer Therapy

November 02-04, 2015 Hyderabad, India

Anuradha Choughule1, Vaishakhi Trivedi1, Pratik Chandrani2, Priyanka Bagayatkar1, Devashree Batham1, Vanita Noronha1, Amit Joshi1, S D Banavali1, Amit Dutt2 and Kumar Prabhash1

1Tata Memorial Hospital, India 2Advanced Centre for Treatment, Research and Education in Cancer, India

Posters-Accepted Abstracts: J Cancer Sci Ther

Abstract :

Introduction & Objectives: Conventional therapeutic solutions in NSCLC are not effective to treat the disease. Despite of all developments in understanding the disease, mortality of lung cancer patients remains high. Recent developments of personalized therapy have given promising results in terms of improved survival of NSCLC patients. Thus, we were keen to develop a cost effective and sensitive diagnostic lung cancer panel assay for targetable mutation detection. Here, we present a multiplexed assay using SNaPshot PCR technique from FFPE samples. Methodology: Multiplex PCR was optimized to amplify hotspot regions from 9 targetable genes followed by single base extension reaction using SNaPshot PCR and fragment analysis on ABI 3500 sequencer. Results: The successfully developed mutation profiling assay was divided into 3 multiplexed reactions, covering 23 actionable genotypes of EGFR, KRAS, BRAF, PIK3CA, Her2, AKT1, NRAS, MEK1 and PTEN genes. The assay was standardized and validated on blood samples, cell lines and FFPE samples expressing good sensitivity and specificity for wild type and mutant genotypes. Conclusion: Multiplexed diagnostic assay using SNaPshot PCR is very economical, specific and sensitive to detect mutations in FFPE samples. We intend to implement this genomic profiling as therapeutic strategy in lung cancer patients at our centre.

Biography :

Email: anu_c1112@hotmail.com