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Sequencing on Exosomal DNA for EGFR Mutation Detection in NSCLC  \nMohammad Mehdi Jahani 1 , Parisa Mashayekhi2* , Mir Davood Omrani 1* , Adnan Khosravi3 , Ali Dehghanifard2 , Sanam Azad Manjiri2 , Mahyar Zahraei2, Maryam Mabani3 , Sharareh Seifi3 , Babak Salimi3 , Parsa Rostami3  \n1Department of Medical Genetics, Shahid Beheshti University of Medical Sciences, Tehran, Iran;  \n2Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran;  \n3Research Center of Thoracic Oncology (RCTO), National Research Institute of Tuberculosis  \nand Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran  \nOPEN ACCESS  \nReceived: June 2, 2024  \nAccepted: June 29, 2024  \nPublished online: June 30, 2024  \nCitation:  \nJahani M, Mashayekhi P, Omrani MD, Khosravi A, Dehghanifard A, Azad Manjiri S, Zahraei M, Mabani M, Seifi S, Salimi B, Rostami P. Assessing the Sensitivity of Nested PCR followed by direct sequencing on Exosomal DNA for EGFR Mutation Detection in NSCLC. Iranian biomedical journal 2024; 28(4): 208-215.  \nABSTRACT  \nBackground: Early and minimally invasive detection of EGFR mutations in NSCLC patients is a promising tool to select patients for targeted therapy in order to improve their prognosis. This study aimed to identify a sensitive, cost-effective, and easily accessible noninvasive method for detecting the EGFR-targetable mutations in the plasma exoDNA of patients with NSCLC. Methods: This retrospective observational study was conducted over 10 months, from December 2022 to October 2023, at Masih Daneshvari Hospital in Tehran, Iran. A total of 30 patients with stage II-IV NSCLC and targetable mutation in the EGFR gene were included in the study. Nested PCR and Sanger sequencing were used to evaluate EGFR mutations in the DNA extracted from circulating exosomes.  \nResults: The study found a sensitivity of 76 .6% for EGFR mutation detection on exoDNA compared to tissue results. No significant impact was observed based on tumor staging, histology, mutation type, smoking status, gender, or age.  \nConclusion: Therapeutically targetable driver mutations in the EGFR gene can be accurately detected using nested PCR followed by direct sequencing of plasma exoDNA from patients with NSCLC. This approach facilitates timely and more personalized treatment for NSCLC patients, ultimately improving patient prognosis. Additionally, this method reduces the reliance on invasive tissue biopsies and their associated complications. DOI: 10.61186/ibj.4289  \nKeywords: Exosomes, Liquid biopsy, Lung neoplasms  \nCorresponding Authors:  \nParisa Mashayekhi  \nMolecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran; Tel.: (+98-11) 22700044;  \nE-mail: [parisamashayekhi7@gmail.com](parisamashayekhi7@gmail.com)  \nMir Davood Omrani  \nUrogenital Stem Cell Research Center, Shahid Labbafinejad Medical Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Tel.: (+98-11) 22567222; E-mail: [davood_omrani@yahoo.co.uk](davood_omrani@yahoo.co.uk)  \nINTRODUCTION  \nAccording to the World Health Organization  \n(2021), lung cancer is a leading cause of cancer  \nrelated deaths, with approximately 12 million incidences of new cases and 1.8 million deaths each year. Among these cases, NSCLC is prevalent in about 85% of individuals. NSCLC is often diagnosed at a later stage, leading to increased therapeutic complexities and lower survival probabilities[1] . Recent advancements in understanding the molecular intricacies of various cancers and the impact of oncogenic driver mutations have paved the way for targeted therapies[2] .  \nEGFR gene mutations are considered targetable drivers in NSCLC, and identifying these mutations has completely altered the approach toward NSCLC treatment[3] . Mutations like exon 19 deletion and also L858R point mutation in exon 21 make patients sensitive to TKIs and","cbCaig1pvmhFfsr7","https://ap.wps.com/l/cbCaig1pvmhFfsr7","pdf",1492232,"English","# Abstract\n## Background and Aim\n## Methods\n## Results\n## Conclusion\n# Introduction\n## Lung cancer burden and need for EGFR testing\n## EGFR mutations and diagnostic limitations of tissue biopsy\n## Rationale for liquid biopsy and exoDNA","[{\"question\":\"What method did the study use to detect EGFR mutations in NSCLC patients?\",\"answer\":\"The study used nested PCR followed by direct sequencing (Sanger sequencing) on DNA extracted from circulating exosomes to detect EGFR mutations in plasma exoDNA.\"},{\"question\":\"How was the exoDNA-based detection performance assessed?\",\"answer\":\"Sensitivity was calculated by comparing EGFR mutation detection on exoDNA with tissue-based results for the included patients.\"},{\"question\":\"What patient-related factors showed no significant effect on the results?\",\"answer\":\"No significant impact was observed by tumor staging, histology, mutation type, smoking status, gender, or age.\"}]","Assessing the Sensitivity of Nested PCR Followed by Direct Sequencing on Exosomal DNA for EGFR Mutation Detection in NSCLC | PDF",1790022749]