Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR

Activating mutations in EGFR predict benefit from tyrosine kinase inhibitor therapy for patients with advanced non-small cell lung cancer. Directing patients to appropriate therapy depends on accurate and timely EGFR assessment in the molecular pathology laboratory. This article describes the analyt...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:PloS one 2022-02, Vol.17 (2), p.e0264201-e0264201
Hauptverfasser: Williamson, Drew F K, Marris, Sean R N, Rojas-Rudilla, Vanesa, Bruce, Jacqueline L, Paweletz, Cloud P, Oxnard, Geoffrey R, Sholl, Lynette M, Dong, Fei
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page e0264201
container_issue 2
container_start_page e0264201
container_title PloS one
container_volume 17
creator Williamson, Drew F K
Marris, Sean R N
Rojas-Rudilla, Vanesa
Bruce, Jacqueline L
Paweletz, Cloud P
Oxnard, Geoffrey R
Sholl, Lynette M
Dong, Fei
description Activating mutations in EGFR predict benefit from tyrosine kinase inhibitor therapy for patients with advanced non-small cell lung cancer. Directing patients to appropriate therapy depends on accurate and timely EGFR assessment in the molecular pathology laboratory. This article describes the analytical design, performance characteristics, and clinical implementation of an assay for the rapid detection of EGFR L858R and exon 19 deletion mutations. A droplet digital polymerase chain reaction (ddPCR) assay was implemented with probe hydrolysis-dependent signal detection. A mutation-specific probe was used to detect EGFR L858R. A loss of signal design was used to detect EGFR exon 19 deletion mutations. Analytical sensitivity was dependent on DNA input and was as low as 0.01% variant allele fraction for the EGFR L858R assay and 0.1% variant allele fraction for the EGFR exon 19 deletion assay. Correlation of 20 clinical specimens tested by ddPCR and next generation sequencing showed 100% concordance. ddPCR showed 53% clinical sensitivity in the detection of EGFR mutations in plasma cell-free DNA from patients with lung cancer. The median clinical turnaround time was 5 days for ddPCR compared to 13 days for next generation sequencing. The findings show that ddPCR is an accurate and rapid method for detecting EGFR mutations in patients with non-small cell lung cancer.
doi_str_mv 10.1371/journal.pone.0264201
format Article
fullrecord <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_2632981771</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A694824556</galeid><doaj_id>oai_doaj_org_article_19219266db6d450e9c5a341aa6bcea1b</doaj_id><sourcerecordid>A694824556</sourcerecordid><originalsourceid>FETCH-LOGICAL-c692t-f8ce493106331eb30629daa07563995f3cf8edbc8724f4ef30151b19732585fb3</originalsourceid><addsrcrecordid>eNqNkl1r2zAUhs3YWLts_2BshsHYLpLpy7J1MyjpxwKFluzjVsiy7CjIUmbJo_33kxO3xKMXw8I2x895j_T6TZK3ECwgzuGXres7K8xi56xaAEQJAvBZcgoZRnOKAH5-9H6SvPJ-C0CGC0pfJic4QwARDE-T23MVlAza2dTV6cXV5Tpt-yCGgk-1Ta2zc98KY1Kp4s30tkmlsFJ1aXmfVp3bGRXSSjc6CJPeLtevkxe1MF69GZ-z5OflxY_lt_n1zdVqeXY9l5ShMK8LqQjDEFCMoSoxoIhVQoA8o5ixrMayLlRVyiJHpCaqxgBmsIQsxygrsrrEs-T9QXdnnOejGZ4jihErYJ7DSKwOROXElu863Yrunjuh-b7guoaLLmhpFIcMxUVpVdKKZEAxmQlMoBC0lErAYdrXcVpftqqSyoZOmIno9IvVG964P7wockAYiwKfRoHO_e6VD7zVfrBUWOX6_b5xQQoA84h--Ad9-nQj1Yh4AG1rF-fKQZSfUUYKRLJo5SxZPEHFq1KtljE5tY71ScPnSUNkgroLjei956vv6_9nb35N2Y9H7EYJEzbemX4ftClIDqDsnPedqh9NhoAPwX9wgw_B52PwY9u74x_02PSQdPwX7tL7YQ</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2632981771</pqid></control><display><type>article</type><title>Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR</title><source>Public Library of Science (PLoS) Journals Open Access</source><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><source>EZB Electronic Journals Library</source><creator>Williamson, Drew F K ; Marris, Sean R N ; Rojas-Rudilla, Vanesa ; Bruce, Jacqueline L ; Paweletz, Cloud P ; Oxnard, Geoffrey R ; Sholl, Lynette M ; Dong, Fei</creator><creatorcontrib>Williamson, Drew F K ; Marris, Sean R N ; Rojas-Rudilla, Vanesa ; Bruce, Jacqueline L ; Paweletz, Cloud P ; Oxnard, Geoffrey R ; Sholl, Lynette M ; Dong, Fei</creatorcontrib><description>Activating mutations in EGFR predict benefit from tyrosine kinase inhibitor therapy for patients with advanced non-small cell lung cancer. Directing patients to appropriate therapy depends on accurate and timely EGFR assessment in the molecular pathology laboratory. This article describes the analytical design, performance characteristics, and clinical implementation of an assay for the rapid detection of EGFR L858R and exon 19 deletion mutations. A droplet digital polymerase chain reaction (ddPCR) assay was implemented with probe hydrolysis-dependent signal detection. A mutation-specific probe was used to detect EGFR L858R. A loss of signal design was used to detect EGFR exon 19 deletion mutations. Analytical sensitivity was dependent on DNA input and was as low as 0.01% variant allele fraction for the EGFR L858R assay and 0.1% variant allele fraction for the EGFR exon 19 deletion assay. Correlation of 20 clinical specimens tested by ddPCR and next generation sequencing showed 100% concordance. ddPCR showed 53% clinical sensitivity in the detection of EGFR mutations in plasma cell-free DNA from patients with lung cancer. The median clinical turnaround time was 5 days for ddPCR compared to 13 days for next generation sequencing. The findings show that ddPCR is an accurate and rapid method for detecting EGFR mutations in patients with non-small cell lung cancer.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0264201</identifier><identifier>PMID: 35202431</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Alleles ; Assaying ; Cancer ; Carcinoma, Non-Small-Cell Lung - genetics ; Deletion ; Deoxyribonucleic acid ; Design ; Diagnosis ; DNA ; DNA Mutational Analysis - methods ; Droplets ; Enzyme inhibitors ; Epidermal growth factor receptors ; ErbB Receptors - genetics ; Flow cytometry ; Gene deletion ; Gene mutations ; Genetic aspects ; Humans ; Kinases ; Laboratories ; Lung cancer ; Lung cancer, Non-small cell ; Lung diseases ; Lung Neoplasms - genetics ; Mutation ; Next-generation sequencing ; Non-small cell lung carcinoma ; Oncology, Experimental ; Pathology ; Patients ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Protein-tyrosine kinase ; Sensitivity analysis ; Sensitivity and Specificity ; Severe acute respiratory syndrome coronavirus 2 ; Signal detection ; Small cell lung carcinoma ; Tyrosine ; Women</subject><ispartof>PloS one, 2022-02, Vol.17 (2), p.e0264201-e0264201</ispartof><rights>COPYRIGHT 2022 Public Library of Science</rights><rights>2022 Williamson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2022 Williamson et al 2022 Williamson et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-f8ce493106331eb30629daa07563995f3cf8edbc8724f4ef30151b19732585fb3</citedby><cites>FETCH-LOGICAL-c692t-f8ce493106331eb30629daa07563995f3cf8edbc8724f4ef30151b19732585fb3</cites><orcidid>0000-0003-3769-0247 ; 0000-0003-1745-8846</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8870499/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8870499/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35202431$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Williamson, Drew F K</creatorcontrib><creatorcontrib>Marris, Sean R N</creatorcontrib><creatorcontrib>Rojas-Rudilla, Vanesa</creatorcontrib><creatorcontrib>Bruce, Jacqueline L</creatorcontrib><creatorcontrib>Paweletz, Cloud P</creatorcontrib><creatorcontrib>Oxnard, Geoffrey R</creatorcontrib><creatorcontrib>Sholl, Lynette M</creatorcontrib><creatorcontrib>Dong, Fei</creatorcontrib><title>Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Activating mutations in EGFR predict benefit from tyrosine kinase inhibitor therapy for patients with advanced non-small cell lung cancer. Directing patients to appropriate therapy depends on accurate and timely EGFR assessment in the molecular pathology laboratory. This article describes the analytical design, performance characteristics, and clinical implementation of an assay for the rapid detection of EGFR L858R and exon 19 deletion mutations. A droplet digital polymerase chain reaction (ddPCR) assay was implemented with probe hydrolysis-dependent signal detection. A mutation-specific probe was used to detect EGFR L858R. A loss of signal design was used to detect EGFR exon 19 deletion mutations. Analytical sensitivity was dependent on DNA input and was as low as 0.01% variant allele fraction for the EGFR L858R assay and 0.1% variant allele fraction for the EGFR exon 19 deletion assay. Correlation of 20 clinical specimens tested by ddPCR and next generation sequencing showed 100% concordance. ddPCR showed 53% clinical sensitivity in the detection of EGFR mutations in plasma cell-free DNA from patients with lung cancer. The median clinical turnaround time was 5 days for ddPCR compared to 13 days for next generation sequencing. The findings show that ddPCR is an accurate and rapid method for detecting EGFR mutations in patients with non-small cell lung cancer.</description><subject>Alleles</subject><subject>Assaying</subject><subject>Cancer</subject><subject>Carcinoma, Non-Small-Cell Lung - genetics</subject><subject>Deletion</subject><subject>Deoxyribonucleic acid</subject><subject>Design</subject><subject>Diagnosis</subject><subject>DNA</subject><subject>DNA Mutational Analysis - methods</subject><subject>Droplets</subject><subject>Enzyme inhibitors</subject><subject>Epidermal growth factor receptors</subject><subject>ErbB Receptors - genetics</subject><subject>Flow cytometry</subject><subject>Gene deletion</subject><subject>Gene mutations</subject><subject>Genetic aspects</subject><subject>Humans</subject><subject>Kinases</subject><subject>Laboratories</subject><subject>Lung cancer</subject><subject>Lung cancer, Non-small cell</subject><subject>Lung diseases</subject><subject>Lung Neoplasms - genetics</subject><subject>Mutation</subject><subject>Next-generation sequencing</subject><subject>Non-small cell lung carcinoma</subject><subject>Oncology, Experimental</subject><subject>Pathology</subject><subject>Patients</subject><subject>Polymerase chain reaction</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Protein-tyrosine kinase</subject><subject>Sensitivity analysis</subject><subject>Sensitivity and Specificity</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><subject>Signal detection</subject><subject>Small cell lung carcinoma</subject><subject>Tyrosine</subject><subject>Women</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNkl1r2zAUhs3YWLts_2BshsHYLpLpy7J1MyjpxwKFluzjVsiy7CjIUmbJo_33kxO3xKMXw8I2x895j_T6TZK3ECwgzuGXres7K8xi56xaAEQJAvBZcgoZRnOKAH5-9H6SvPJ-C0CGC0pfJic4QwARDE-T23MVlAza2dTV6cXV5Tpt-yCGgk-1Ta2zc98KY1Kp4s30tkmlsFJ1aXmfVp3bGRXSSjc6CJPeLtevkxe1MF69GZ-z5OflxY_lt_n1zdVqeXY9l5ShMK8LqQjDEFCMoSoxoIhVQoA8o5ixrMayLlRVyiJHpCaqxgBmsIQsxygrsrrEs-T9QXdnnOejGZ4jihErYJ7DSKwOROXElu863Yrunjuh-b7guoaLLmhpFIcMxUVpVdKKZEAxmQlMoBC0lErAYdrXcVpftqqSyoZOmIno9IvVG964P7wockAYiwKfRoHO_e6VD7zVfrBUWOX6_b5xQQoA84h--Ad9-nQj1Yh4AG1rF-fKQZSfUUYKRLJo5SxZPEHFq1KtljE5tY71ScPnSUNkgroLjei956vv6_9nb35N2Y9H7EYJEzbemX4ftClIDqDsnPedqh9NhoAPwX9wgw_B52PwY9u74x_02PSQdPwX7tL7YQ</recordid><startdate>20220224</startdate><enddate>20220224</enddate><creator>Williamson, Drew F K</creator><creator>Marris, Sean R N</creator><creator>Rojas-Rudilla, Vanesa</creator><creator>Bruce, Jacqueline L</creator><creator>Paweletz, Cloud P</creator><creator>Oxnard, Geoffrey R</creator><creator>Sholl, Lynette M</creator><creator>Dong, Fei</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0003-3769-0247</orcidid><orcidid>https://orcid.org/0000-0003-1745-8846</orcidid></search><sort><creationdate>20220224</creationdate><title>Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR</title><author>Williamson, Drew F K ; Marris, Sean R N ; Rojas-Rudilla, Vanesa ; Bruce, Jacqueline L ; Paweletz, Cloud P ; Oxnard, Geoffrey R ; Sholl, Lynette M ; Dong, Fei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-f8ce493106331eb30629daa07563995f3cf8edbc8724f4ef30151b19732585fb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Alleles</topic><topic>Assaying</topic><topic>Cancer</topic><topic>Carcinoma, Non-Small-Cell Lung - genetics</topic><topic>Deletion</topic><topic>Deoxyribonucleic acid</topic><topic>Design</topic><topic>Diagnosis</topic><topic>DNA</topic><topic>DNA Mutational Analysis - methods</topic><topic>Droplets</topic><topic>Enzyme inhibitors</topic><topic>Epidermal growth factor receptors</topic><topic>ErbB Receptors - genetics</topic><topic>Flow cytometry</topic><topic>Gene deletion</topic><topic>Gene mutations</topic><topic>Genetic aspects</topic><topic>Humans</topic><topic>Kinases</topic><topic>Laboratories</topic><topic>Lung cancer</topic><topic>Lung cancer, Non-small cell</topic><topic>Lung diseases</topic><topic>Lung Neoplasms - genetics</topic><topic>Mutation</topic><topic>Next-generation sequencing</topic><topic>Non-small cell lung carcinoma</topic><topic>Oncology, Experimental</topic><topic>Pathology</topic><topic>Patients</topic><topic>Polymerase chain reaction</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Protein-tyrosine kinase</topic><topic>Sensitivity analysis</topic><topic>Sensitivity and Specificity</topic><topic>Severe acute respiratory syndrome coronavirus 2</topic><topic>Signal detection</topic><topic>Small cell lung carcinoma</topic><topic>Tyrosine</topic><topic>Women</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Williamson, Drew F K</creatorcontrib><creatorcontrib>Marris, Sean R N</creatorcontrib><creatorcontrib>Rojas-Rudilla, Vanesa</creatorcontrib><creatorcontrib>Bruce, Jacqueline L</creatorcontrib><creatorcontrib>Paweletz, Cloud P</creatorcontrib><creatorcontrib>Oxnard, Geoffrey R</creatorcontrib><creatorcontrib>Sholl, Lynette M</creatorcontrib><creatorcontrib>Dong, Fei</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale in Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing &amp; Allied Health Database (ProQuest)</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological &amp; Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health &amp; Medical Collection (Proquest)</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database (Proquest)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science &amp; Engineering Database (Proquest)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies &amp; Aerospace Collection</collection><collection>ProQuest Agriculture &amp; Environmental Science Database</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>ProQuest Materials Science Database</collection><collection>Nursing &amp; Allied Health Database (Alumni Edition)</collection><collection>Meteorological &amp; Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>Biological Sciences</collection><collection>Agriculture Science Database</collection><collection>Health &amp; Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>ProQuest Engineering Database</collection><collection>Nursing &amp; Allied Health Premium</collection><collection>ProQuest advanced technologies &amp; aerospace journals</collection><collection>ProQuest Advanced Technologies &amp; Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Engineering collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Williamson, Drew F K</au><au>Marris, Sean R N</au><au>Rojas-Rudilla, Vanesa</au><au>Bruce, Jacqueline L</au><au>Paweletz, Cloud P</au><au>Oxnard, Geoffrey R</au><au>Sholl, Lynette M</au><au>Dong, Fei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2022-02-24</date><risdate>2022</risdate><volume>17</volume><issue>2</issue><spage>e0264201</spage><epage>e0264201</epage><pages>e0264201-e0264201</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Activating mutations in EGFR predict benefit from tyrosine kinase inhibitor therapy for patients with advanced non-small cell lung cancer. Directing patients to appropriate therapy depends on accurate and timely EGFR assessment in the molecular pathology laboratory. This article describes the analytical design, performance characteristics, and clinical implementation of an assay for the rapid detection of EGFR L858R and exon 19 deletion mutations. A droplet digital polymerase chain reaction (ddPCR) assay was implemented with probe hydrolysis-dependent signal detection. A mutation-specific probe was used to detect EGFR L858R. A loss of signal design was used to detect EGFR exon 19 deletion mutations. Analytical sensitivity was dependent on DNA input and was as low as 0.01% variant allele fraction for the EGFR L858R assay and 0.1% variant allele fraction for the EGFR exon 19 deletion assay. Correlation of 20 clinical specimens tested by ddPCR and next generation sequencing showed 100% concordance. ddPCR showed 53% clinical sensitivity in the detection of EGFR mutations in plasma cell-free DNA from patients with lung cancer. The median clinical turnaround time was 5 days for ddPCR compared to 13 days for next generation sequencing. The findings show that ddPCR is an accurate and rapid method for detecting EGFR mutations in patients with non-small cell lung cancer.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>35202431</pmid><doi>10.1371/journal.pone.0264201</doi><tpages>e0264201</tpages><orcidid>https://orcid.org/0000-0003-3769-0247</orcidid><orcidid>https://orcid.org/0000-0003-1745-8846</orcidid><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 1932-6203
ispartof PloS one, 2022-02, Vol.17 (2), p.e0264201-e0264201
issn 1932-6203
1932-6203
language eng
recordid cdi_plos_journals_2632981771
source Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; PubMed Central; Free Full-Text Journals in Chemistry; EZB Electronic Journals Library
subjects Alleles
Assaying
Cancer
Carcinoma, Non-Small-Cell Lung - genetics
Deletion
Deoxyribonucleic acid
Design
Diagnosis
DNA
DNA Mutational Analysis - methods
Droplets
Enzyme inhibitors
Epidermal growth factor receptors
ErbB Receptors - genetics
Flow cytometry
Gene deletion
Gene mutations
Genetic aspects
Humans
Kinases
Laboratories
Lung cancer
Lung cancer, Non-small cell
Lung diseases
Lung Neoplasms - genetics
Mutation
Next-generation sequencing
Non-small cell lung carcinoma
Oncology, Experimental
Pathology
Patients
Polymerase chain reaction
Polymerase Chain Reaction - methods
Protein-tyrosine kinase
Sensitivity analysis
Sensitivity and Specificity
Severe acute respiratory syndrome coronavirus 2
Signal detection
Small cell lung carcinoma
Tyrosine
Women
title Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-01T11%3A00%3A14IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Detection%20of%20EGFR%20mutations%20in%20non-small%20cell%20lung%20cancer%20by%20droplet%20digital%20PCR&rft.jtitle=PloS%20one&rft.au=Williamson,%20Drew%20F%20K&rft.date=2022-02-24&rft.volume=17&rft.issue=2&rft.spage=e0264201&rft.epage=e0264201&rft.pages=e0264201-e0264201&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0264201&rft_dat=%3Cgale_plos_%3EA694824556%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2632981771&rft_id=info:pmid/35202431&rft_galeid=A694824556&rft_doaj_id=oai_doaj_org_article_19219266db6d450e9c5a341aa6bcea1b&rfr_iscdi=true