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A peptide nucleic acid (PNA) probe-based mutant mRNA detection approach offers rapid and low-cost testing. This study evaluates PNA:DNA probe efficiency for detecting EGFR mutations in formalin-fixed paraffin-embedded (FFPE) NSCLC patient specimens and examines how EGFR mutation localization relates to tumor-infiltrating lymphocyte (TIL) status.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/localization-of-epidermal-growth-factor-receptor-mutations-using-pnadna-probes-in-clinical-specimens-from-patients-with-nonsmall-cell-lung-cancer/372262/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/localization-of-epidermal-growth-factor-receptor-mutations-using-pnadna-probes-in-clinical-specimens-from-patients-with-nonsmall-cell-lung-cancer/372262.png","ImageObject",300,407,{"name":92,"@type":93},"Genevieve","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-29","2026-09-23",true,{"@type":102,"interactionType":103,"userInteractionCount":44},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"Why is the study needed instead of relying on NGS-based mutation detection?","Question",{"text":112,"@type":113},"NGS mutation detection is accurate but time-consuming and expensive. The study targets faster and lower-cost mutation detection using PNA:DNA probes.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How were EGFR mutations detected in the specimens?",{"text":117,"@type":113},"The work used EGFR mutation-specific PNA:DNA probes to detect EGFR mutations in FFPE specimens, including the L858R-specific probe.",{"name":119,"@type":110,"acceptedAnswer":120},"What does the study report about tumor-infiltrating lymphocytes (TILs) in relation to EGFR mutations?",{"text":121,"@type":113},"TIL analysis showed scarce CD8+PD-1+ T cells and CD68+ macrophages in L858R-mutated tumors overall, while CD4+, FoxP3+, and CD204+ cells tended to be more abundant in cytokeratin-positive intra-tumoral regions of L858R-positive areas.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},372262,1790621195,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":44,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":52,"language":139,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":140,"faqs":141,"seo_title":142,"seo_description":67,"update_tm":143,"read_time":144},1374391974585,"https://ap-avatar.wpscdn.com/davatar_276721f389ce27ea32af1340a28f341c","[www. nature.com/scientificreports](www. nature.com/scientificreports)  \nOPEN  \nLocalization of epidermal growth factor receptor-mutations using PNA:DNA probes in clinical specimens from patients with nonsmall cell lung cancer  \nHaruo Miyata1,12, Hajime Shigeto2,12, Tomoatsu Ikeya1, Tadashi Ashizawa1, Akira Iizuka1, Yasufumi Kikuchi1, Chie Maeda1, Akari Kanematsu1, Kazue Yamashita1, Kenichi Urakami3, Yuji Shimoda3, Takeshi Nagashima3,4, Keiichi Ohshima5, Yasuhisa Ohde6, Mitsuhiro Isaka6, Takashi Sugino7, Koji Muramatsu7, Hirotsugu Kenmotsu8, Toshiaki Takahashi9, Ken Yamaguchi10, Mizuki Kitamatsu11, Shohei Yamamura2 & YasutoAkiyama1􀀍  \nQuickly identifying driver gene mutations in solid cancers is important. However, next-generation sequencing (NGS)-based mutation detection methods are time-consuming and expensive. Peptide nucleic acid (PNA) probe-based mutant mRNA detection systems are quick and inexpensive. We previously demonstrated that epidermal growth factor receptor (EGFR)-mutations were efficiently visualized in formalin-fixed paraffin-embedded (FFPE) specimens from transplanted non-small cell lung cancer (NSCLC) tumors using an EGFR mutation-specific PNA:DNA probe. Herein, the efficiency of PNA:DNA probes in detecting EGFR-mutations in FFPE specimens from patients with NSCLC and the colocalization of EGFR-mutations with tumor-infiltrating lymphocyte (TIL) status were determined. The EGFR mutation L858R-specific PNA:DNA probe detected heterogeneously localized mutations with a sensitivity similar to detection with the anti-L858R antibody. TIL analysis of L858Rmutated tumors revealed that CD8+PD-1+ T cells and CD68+ macrophages were scarce in tumors, but in the cytokeratin-positive intra-tumoral regions, CD4+, FoxP3+, and CD204+ cells tended to be more abundant inthe L858R-positive tumor area than in the negative area. Thus, PNA:DNA probes specific for EGFR-mutations can detect areas with heterogeneous EGFR mutants in whole cancer tissues and can be used to evaluate the mutation-associated TIL status in EGFR-mutant cancer tissues.  \nKeywords Cancer genome analysis, Epidermal growth factor receptor mutation, Next generation sequencing, PNA probe, Tumor-infiltrating lymphocyte (TIL)  \nA peptide nucleic acid (PNA)-based mutant mRNA detection system was developed by Nielsen et al., prompting many trials applying PNA probes to the molecular genetics field1–5. In addition to the speed and low-cost, many advantages ofthe PNA technology have been demonstrated, including high sensitivity and specificity for hybridization to complementary DNA or RNA, single base mismatch discrimination, highly stable binding to DNA or RNA due to the higher melting temperature, and resistance to nuclease digestion and low ionic strength conditions due to the uncharged backbone6–8.  \n1Immunotherapy Division, Shizuoka Cancer Center Research Institute, 1007 Shimonagakubo, Nagaizumi-cho, Sunto-gun, Shizuoka 411-8777, Japan. 2Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Kagawa, Japan. 3Cancer Diagnostics Research Division, Shizuoka Cancer Center Research Institute, Shizuoka, Japan. 4SRL Inc., Tokyo, Japan. 5Medical Genetics Division, Shizuoka Cancer Center Research Institute, Shizuoka, Japan. 6Division of Thoracic Surgery, Shizuoka Cancer Center Hospital, Shizuoka, Japan. 7Division of Pathology, Shizuoka Cancer Center Hospital, Shizuoka, Japan. 8Division of Genetic Medicine Promotion, Shizuoka Cancer Center Hospital, Shizuoka, Japan. 9Division of Thoracic Oncology, Shizuoka Cancer Center Hospital, Shizuoka, Japan. 10Office of the President Emeritus, Shizuoka Cancer Center Hospital, Shizuoka, Japan. 11Department of Applied Chemistry, Kindai University, Osaka, Japan. 12Haruo Miyata and Hajime Shigeto contributed equally to this work. 􀀍 [email: y.akiyama@scchr.jp](email: y.akiyama@scchr.jp)  \n[www. nature.com/scientificreports/](www. nature.com/scientificreports/)  \n\n| EGFR mutations | Case ","cbCaipmsLS4qRF8Z","https://ap.wps.com/l/cbCaipmsLS4qRF8Z","pdf",3006674,"English","# Localization and detection approach\n## Study objectives and rationale\n## Probe performance in FFPE specimens\n## EGFR mutation localization and TIL correlations","[{\"question\":\"Why is the study needed instead of relying on NGS-based mutation detection?\",\"answer\":\"NGS mutation detection is accurate but time-consuming and expensive. The study targets faster and lower-cost mutation detection using PNA:DNA probes.\"},{\"question\":\"How were EGFR mutations detected in the specimens?\",\"answer\":\"The work used EGFR mutation-specific PNA:DNA probes to detect EGFR mutations in FFPE specimens, including the L858R-specific probe.\"},{\"question\":\"What does the study report about tumor-infiltrating lymphocytes (TILs) in relation to EGFR mutations?\",\"answer\":\"TIL analysis showed scarce CD8+PD-1+ T cells and CD68+ macrophages in L858R-mutated tumors overall, while CD4+, FoxP3+, and CD204+ cells tended to be more abundant in cytokeratin-positive intra-tumoral regions of L858R-positive areas.\"}]","Localization of epidermal growth factor receptor-mutations using PNA:DNA probes in clinical specimens from patients with nonsmall cell lung cancer | PDF",1790194102,25]