Screening for EGFR and KRAS Mutations in Endobronchial Ultrasound Derived Transbronchial Needle Aspirates in Non-Small Cell Lung Cancer Using COLD-PCR
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{"title"=>"Screening for EGFR and KRAS mutations in endobronchial ultrasound derived transbronchial needle aspirates in non-small cell lung cancer using COLD-PCR", "type"=>"journal", "authors"=>[{"first_name"=>"George", "last_name"=>"Santis", "scopus_author_id"=>"6701907274"}, {"first_name"=>"Roger", "last_name"=>"Angell", "scopus_author_id"=>"33067506100"}, {"first_name"=>"Guillermina", "last_name"=>"Nickless", "scopus_author_id"=>"37261692700"}, {"first_name"=>"Alison", "last_name"=>"Quinn", "scopus_author_id"=>"52664390700"}, {"first_name"=>"Amanda", "last_name"=>"Herbert", "scopus_author_id"=>"7103273544"}, {"first_name"=>"Paul", "last_name"=>"Cane", "scopus_author_id"=>"24477271400"}, {"first_name"=>"James", "last_name"=>"Spicer", "scopus_author_id"=>"7103113712"}, {"first_name"=>"Ronan", "last_name"=>"Breen", "scopus_author_id"=>"8935673400"}, {"first_name"=>"Emma", "last_name"=>"McLean", "scopus_author_id"=>"36559876100"}, {"first_name"=>"Khalid", "last_name"=>"Tobal", "scopus_author_id"=>"6701713469"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-80052892102", "doi"=>"10.1371/journal.pone.0025191", "sgr"=>"80052892102", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "pmid"=>"21949883", "issn"=>"19326203", "pui"=>"362577515"}, "id"=>"021eb533-c744-38d9-9495-ea8e4f9cb9ab", "abstract"=>"EGFR mutations correlate with improved clinical outcome whereas KRAS mutations are associated with lack of response to tyrosine kinase inhibitors in patients with non-small cell lung cancer (NSCLC). Endobronchial ultrasound (EBUS)-transbronchial needle aspiration (TBNA) is being increasingly used in the management of NSCLC. Co-amplification at lower denaturation temperature (COLD)-polymerase chain reaction (PCR) (COLD-PCR) is a sensitive assay for the detection of genetic mutations in solid tumours. This study assessed the feasibility of using COLD-PCR to screen for EGFR and KRAS mutations in cytology samples obtained by EBUS-TBNA in routine clinical practice. Samples obtained from NSCLC patients undergoing EBUS-TBNA were evaluated according to our standard clinical protocols. DNA extracted from these samples was subjected to COLD-PCR to amplify exons 18-21 of EGFR and exons two and three of KRAS followed by direct sequencing. Mutation analysis was performed in 131 of 132 (99.3%) NSCLC patients (70F/62M) with confirmed lymph node metastases (94/132 (71.2%) adenocarcinoma; 17/132 (12.8%) squamous cell; 2/132 (0.15%) large cell neuroendocrine; 1/132 (0.07%) large cell carcinoma; 18/132 (13.6%) NSCL-not otherwise specified (NOS)). Molecular analysis of all EGFR and KRAS target sequences was achieved in 126 of 132 (95.5%) and 130 of 132 (98.4%) of cases respectively. EGFR mutations were identified in 13 (10.5%) of fully evaluated cases (11 in adenocarcinoma and two in NSCLC-NOS) including two novel mutations. KRAS mutations were identified in 23 (17.5%) of fully analysed patient samples (18 adenocarcinoma and five NSCLC-NOS). We conclude that EBUS-TBNA of lymph nodes infiltrated by NSCLC can provide sufficient tumour material for EGFR and KRAS mutation analysis in most patients, and that COLD-PCR and sequencing is a robust screening assay for EGFR and KRAS mutation analysis in this clinical context.", "link"=>"http://www.mendeley.com/research/screening-egfr-kras-mutations-endobronchial-ultrasound-derived-transbronchial-needle-aspirates-nonsm-3", "reader_count"=>33, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Librarian"=>1, "Student > Doctoral Student"=>2, "Researcher"=>10, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>2, "Other"=>4, "Student > Master"=>1, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Librarian"=>1, "Student > Doctoral Student"=>2, "Researcher"=>10, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>2, "Other"=>4, "Student > Master"=>1, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>3, "Medicine and Dentistry"=>18, "Agricultural and Biological Sciences"=>9}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>18}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Netherlands"=>1, "United States"=>1, "Japan"=>1}, "group_count"=>3}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/734976"], "description"=>"<p>Sequencing electrogramme of comparative analysis of COLD-PCR and standard-PCR amplification of <i>EGFR</i> exon 19 and <i>KRAS</i> exon 2. A: upper and lower panels are COLD and standard PCR amplification of exon 21 of <i>EGFR</i> from EBUS-derived aspirates from lymph nodes infiltrated by metastatic lung adenocarcinoma respectively. A shows substitution of thymidine (T) by cytosine (C) in exon 21 of <i>EGFR</i> to generate L858R mutation that is evident in the COLD-PCR amplification reaction (arrow) as well as the standard PCR reaction. The mutation peak is more clearly visible in the COLD-PCR (upper panel) compared to standard-PCR reaction (lower panel). B upper and lower panels are COLD and standard PCR amplification of <i>KRAS</i> exon 2. The upper panel shows substitution of guanine (C) by thymidine (T) to generate G12C mutation that was detected by COLD-PCR amplification (arrow). The mutation was not evident in the standard-PCR reaction (lower panel).</p>", "links"=>[], "tags"=>["comparative", "cold-pcr"], "article_id"=>405312, "categories"=>["Cancer", "Genetics"], "users"=>["George Santis", "Roger Angell", "Guillermina Nickless", "Alison Quinn", "Amanda Herbert", "Paul Cane", "James Spicer", "Ronan Breen", "Emma McLean", "Khalid Tobal"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025191.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Examples_of_comparative_analysis_of_COLD_PCR_vs_standard_PCR_/405312", "title"=>"Examples of comparative analysis of COLD-PCR <i>vs</i> standard-PCR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:43:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/735115"], "description"=>"<p>*Refers to not otherwise specified.</p>+<p>Refers to delineation of lymph node stations by endobronchial ultrasound (EBUS) was based on the new International Association of Study of Lung Cancer (IASLC) lymph node map.</p>", "links"=>[], "tags"=>["genetics and genomics", "oncology"], "article_id"=>405461, "categories"=>["Cancer", "Genetics"], "users"=>["George Santis", "Roger Angell", "Guillermina Nickless", "Alison Quinn", "Amanda Herbert", "Paul Cane", "James Spicer", "Ronan Breen", "Emma McLean", "Khalid Tobal"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025191.t001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Patient_Characteristics_/405461", "title"=>"Patient Characteristics.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 14:44:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/735145"], "description"=>"<p>PCR Oligonucleotide Sequences.</p>", "links"=>[], "tags"=>["oligonucleotide"], "article_id"=>405500, "categories"=>["Cancer", "Genetics"], "users"=>["George Santis", "Roger Angell", "Guillermina Nickless", "Alison Quinn", "Amanda Herbert", "Paul Cane", "James Spicer", "Ronan Breen", "Emma McLean", "Khalid Tobal"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025191.t004", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCR_Oligonucleotide_Sequences_/405500", "title"=>"PCR Oligonucleotide Sequences.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 14:44:22"}
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  • {"files"=>["https://ndownloader.figshare.com/files/735037"], "description"=>"<p>*Refers to data for fully analysed patient samples.</p>+<p>Refers to not otherwise specified.</p>", "links"=>[], "tags"=>["mutations", "metastatic", "lymph", "nodes"], "article_id"=>405386, "categories"=>["Cancer", "Genetics"], "users"=>["George Santis", "Roger Angell", "Guillermina Nickless", "Alison Quinn", "Amanda Herbert", "Paul Cane", "James Spicer", "Ronan Breen", "Emma McLean", "Khalid Tobal"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025191.t002", "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Frequency_of_EGFR_and_KRAS_mutations_in_metastatic_lymph_nodes_in_NSCLC_/405386", "title"=>"Frequency of <i>EGFR</i> and <i>KRAS</i> mutations in metastatic lymph nodes in NSCLC<sup>*</sup>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 14:43:44"}

PMC Usage Stats | Further Information

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Relative Metric

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