Comparison of digital PCR systems for the analysis of liquid biopsy samples of patients affected by lung and colorectal cancer
•Digital PCR is useful for tumor molecular characterization via liquid biopsy.•A moderate agreement between droplet and solid digital PCR was observed.•Solid digital PCR has a higher sensitivity in the detection of mutated cases. Highly sensitive technologies are available for the molecular characte...
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Veröffentlicht in: | Clinica chimica acta 2023-02, Vol.541, p.117239-117239, Article 117239 |
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creator | Crucitta, Stefania Ruglioni, Martina Novi, Claudia Manganiello, Mascia Arici, Roberta Petrini, Iacopo Pardini, Eleonora Cucchiara, Federico Marmorino, Federica Cremolini, Chiara Fogli, Stefano Danesi, Romano Del Re, Marzia |
description | •Digital PCR is useful for tumor molecular characterization via liquid biopsy.•A moderate agreement between droplet and solid digital PCR was observed.•Solid digital PCR has a higher sensitivity in the detection of mutated cases.
Highly sensitive technologies are available for the molecular characterization of solid tumors, including digital PCR (dPCR). Liquid biopsy, based on the analysis of cell-free DNA (cfDNA), is often used to assess EGFR or RAS alterations in lung and colorectal cancers. Our study aimed to compare the results of two different dPCR platforms for the detection of mutations in cfDNA.
Plasma samples from lung and colorectal cancer patients collected as per routine procedures have been tested. cfDNA Was extracted from plasma, and samples were screened on the droplet digital PCR (ddPCR, BioRad) and solid dPCR QIAcuity (Qiagen).
A total of 42 samples were analyzed, obtained from 20 Non-Small Cell Lung Cancer (NSCLC) patients carrying an EGFR or a KRAS mutation on tissue at diagnosis, and from 22 samples of colorectal cancer (CRC) patients, 10 of which presenting a KRAS mutation. EGFR mutation detection was 58.8% for ddPCR and 100% for dPCR (κ = 0.54; 95% CI, 0.37–0.71), compared to tissue results. The detection rate for RAS mutations was 72.7% for ddPCR and 86.4% for dPCR (κ = 0.34; 95% CI, 0.01–0.68), compared to tissue results.
This study showed moderate agreement between dPCR and ddPCR. Sampling effect or threshold settings may potentially explain the differences in the cfDNA data between the two different platforms. |
doi_str_mv | 10.1016/j.cca.2023.117239 |
format | Article |
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Highly sensitive technologies are available for the molecular characterization of solid tumors, including digital PCR (dPCR). Liquid biopsy, based on the analysis of cell-free DNA (cfDNA), is often used to assess EGFR or RAS alterations in lung and colorectal cancers. Our study aimed to compare the results of two different dPCR platforms for the detection of mutations in cfDNA.
Plasma samples from lung and colorectal cancer patients collected as per routine procedures have been tested. cfDNA Was extracted from plasma, and samples were screened on the droplet digital PCR (ddPCR, BioRad) and solid dPCR QIAcuity (Qiagen).
A total of 42 samples were analyzed, obtained from 20 Non-Small Cell Lung Cancer (NSCLC) patients carrying an EGFR or a KRAS mutation on tissue at diagnosis, and from 22 samples of colorectal cancer (CRC) patients, 10 of which presenting a KRAS mutation. EGFR mutation detection was 58.8% for ddPCR and 100% for dPCR (κ = 0.54; 95% CI, 0.37–0.71), compared to tissue results. The detection rate for RAS mutations was 72.7% for ddPCR and 86.4% for dPCR (κ = 0.34; 95% CI, 0.01–0.68), compared to tissue results.
This study showed moderate agreement between dPCR and ddPCR. Sampling effect or threshold settings may potentially explain the differences in the cfDNA data between the two different platforms.</description><identifier>ISSN: 0009-8981</identifier><identifier>EISSN: 1873-3492</identifier><identifier>DOI: 10.1016/j.cca.2023.117239</identifier><identifier>PMID: 36736684</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Carcinoma, Non-Small-Cell Lung ; Cell-Free Nucleic Acids ; cfDNA ; Colorectal cancer ; Colorectal Neoplasms - genetics ; Digital PCR ; ErbB Receptors - genetics ; Humans ; Liquid Biopsy ; Lung ; Lung - pathology ; Lung Neoplasms - diagnosis ; Mutation ; Polymerase Chain Reaction - methods ; Proto-Oncogene Proteins p21(ras) - genetics</subject><ispartof>Clinica chimica acta, 2023-02, Vol.541, p.117239-117239, Article 117239</ispartof><rights>2023 The Authors</rights><rights>Copyright © 2023 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c396t-746de3de7fa3845c27db2cf28a089da0dc7989c18eb4d531b4b32991226e3b513</citedby><cites>FETCH-LOGICAL-c396t-746de3de7fa3845c27db2cf28a089da0dc7989c18eb4d531b4b32991226e3b513</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.cca.2023.117239$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36736684$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Crucitta, Stefania</creatorcontrib><creatorcontrib>Ruglioni, Martina</creatorcontrib><creatorcontrib>Novi, Claudia</creatorcontrib><creatorcontrib>Manganiello, Mascia</creatorcontrib><creatorcontrib>Arici, Roberta</creatorcontrib><creatorcontrib>Petrini, Iacopo</creatorcontrib><creatorcontrib>Pardini, Eleonora</creatorcontrib><creatorcontrib>Cucchiara, Federico</creatorcontrib><creatorcontrib>Marmorino, Federica</creatorcontrib><creatorcontrib>Cremolini, Chiara</creatorcontrib><creatorcontrib>Fogli, Stefano</creatorcontrib><creatorcontrib>Danesi, Romano</creatorcontrib><creatorcontrib>Del Re, Marzia</creatorcontrib><title>Comparison of digital PCR systems for the analysis of liquid biopsy samples of patients affected by lung and colorectal cancer</title><title>Clinica chimica acta</title><addtitle>Clin Chim Acta</addtitle><description>•Digital PCR is useful for tumor molecular characterization via liquid biopsy.•A moderate agreement between droplet and solid digital PCR was observed.•Solid digital PCR has a higher sensitivity in the detection of mutated cases.
Highly sensitive technologies are available for the molecular characterization of solid tumors, including digital PCR (dPCR). Liquid biopsy, based on the analysis of cell-free DNA (cfDNA), is often used to assess EGFR or RAS alterations in lung and colorectal cancers. Our study aimed to compare the results of two different dPCR platforms for the detection of mutations in cfDNA.
Plasma samples from lung and colorectal cancer patients collected as per routine procedures have been tested. cfDNA Was extracted from plasma, and samples were screened on the droplet digital PCR (ddPCR, BioRad) and solid dPCR QIAcuity (Qiagen).
A total of 42 samples were analyzed, obtained from 20 Non-Small Cell Lung Cancer (NSCLC) patients carrying an EGFR or a KRAS mutation on tissue at diagnosis, and from 22 samples of colorectal cancer (CRC) patients, 10 of which presenting a KRAS mutation. EGFR mutation detection was 58.8% for ddPCR and 100% for dPCR (κ = 0.54; 95% CI, 0.37–0.71), compared to tissue results. The detection rate for RAS mutations was 72.7% for ddPCR and 86.4% for dPCR (κ = 0.34; 95% CI, 0.01–0.68), compared to tissue results.
This study showed moderate agreement between dPCR and ddPCR. Sampling effect or threshold settings may potentially explain the differences in the cfDNA data between the two different platforms.</description><subject>Carcinoma, Non-Small-Cell Lung</subject><subject>Cell-Free Nucleic Acids</subject><subject>cfDNA</subject><subject>Colorectal cancer</subject><subject>Colorectal Neoplasms - genetics</subject><subject>Digital PCR</subject><subject>ErbB Receptors - genetics</subject><subject>Humans</subject><subject>Liquid Biopsy</subject><subject>Lung</subject><subject>Lung - pathology</subject><subject>Lung Neoplasms - diagnosis</subject><subject>Mutation</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Proto-Oncogene Proteins p21(ras) - genetics</subject><issn>0009-8981</issn><issn>1873-3492</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kEuLFDEURoMoTjv6A9xIlm6qzavzwJU04wMGFNF1SCW3xjSpSk1SJdTG327aHl26Cjf3fF_IQeglJXtKqHxz2nvv9owwvqdUMW4eoR3VindcGPYY7QghptNG0yv0rNZTGwWR9Cm64lJxKbXYoV_HPM6uxJonnAcc4l1cXMJfjl9x3eoCY8VDLnj5AdhNLm011jOX4v0aA-5jnuuGqxvnBH8Ws1siTEvFbhjAL9CYDad1umvxgH1OubTr9oJ3k4fyHD0ZXKrw4uG8Rt_f33w7fuxuP3_4dHx323lu5NIpIQPwAGpwXIuDZyr0zA9MO6JNcCR4ZbTxVEMvwoHTXvScGUMZk8D7A-XX6PWldy75foW62DFWDym5CfJaLVOKUyqJEA2lF9SXXGuBwc4ljq5slhJ71m5Ptmm3Z-32or1lXj3Ur_0I4V_ir-cGvL0A0D75M0Kx1TdPHkI8-7Ahx__U_waqoJP4</recordid><startdate>20230215</startdate><enddate>20230215</enddate><creator>Crucitta, Stefania</creator><creator>Ruglioni, Martina</creator><creator>Novi, Claudia</creator><creator>Manganiello, Mascia</creator><creator>Arici, Roberta</creator><creator>Petrini, Iacopo</creator><creator>Pardini, Eleonora</creator><creator>Cucchiara, Federico</creator><creator>Marmorino, Federica</creator><creator>Cremolini, Chiara</creator><creator>Fogli, Stefano</creator><creator>Danesi, Romano</creator><creator>Del Re, Marzia</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</scope><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>7X8</scope></search><sort><creationdate>20230215</creationdate><title>Comparison of digital PCR systems for the analysis of liquid biopsy samples of patients affected by lung and colorectal cancer</title><author>Crucitta, Stefania ; 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Highly sensitive technologies are available for the molecular characterization of solid tumors, including digital PCR (dPCR). Liquid biopsy, based on the analysis of cell-free DNA (cfDNA), is often used to assess EGFR or RAS alterations in lung and colorectal cancers. Our study aimed to compare the results of two different dPCR platforms for the detection of mutations in cfDNA.
Plasma samples from lung and colorectal cancer patients collected as per routine procedures have been tested. cfDNA Was extracted from plasma, and samples were screened on the droplet digital PCR (ddPCR, BioRad) and solid dPCR QIAcuity (Qiagen).
A total of 42 samples were analyzed, obtained from 20 Non-Small Cell Lung Cancer (NSCLC) patients carrying an EGFR or a KRAS mutation on tissue at diagnosis, and from 22 samples of colorectal cancer (CRC) patients, 10 of which presenting a KRAS mutation. EGFR mutation detection was 58.8% for ddPCR and 100% for dPCR (κ = 0.54; 95% CI, 0.37–0.71), compared to tissue results. The detection rate for RAS mutations was 72.7% for ddPCR and 86.4% for dPCR (κ = 0.34; 95% CI, 0.01–0.68), compared to tissue results.
This study showed moderate agreement between dPCR and ddPCR. Sampling effect or threshold settings may potentially explain the differences in the cfDNA data between the two different platforms.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>36736684</pmid><doi>10.1016/j.cca.2023.117239</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Carcinoma, Non-Small-Cell Lung Cell-Free Nucleic Acids cfDNA Colorectal cancer Colorectal Neoplasms - genetics Digital PCR ErbB Receptors - genetics Humans Liquid Biopsy Lung Lung - pathology Lung Neoplasms - diagnosis Mutation Polymerase Chain Reaction - methods Proto-Oncogene Proteins p21(ras) - genetics |
title | Comparison of digital PCR systems for the analysis of liquid biopsy samples of patients affected by lung and colorectal cancer |
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