Analytical Performance of a qRT-PCR Assay to Detect Guanylyl Cyclase C in FFPE Lymph Nodes of Patients With Colon Cancer
Up to 30% of patients with stage II (pN0) colon cancer develop recurrences, suggesting that the presence of lymph node (LN) metastases escaped detection at histopathologic staging. A simple way to overcome this limitation and to improve staging accuracy is to use reverse transcription-polymerase cha...
Gespeichert in:
| Veröffentlicht in: | Diagnostic molecular pathology 2010-03, Vol.19 (1), p.20-27 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 27 |
|---|---|
| container_issue | 1 |
| container_start_page | 20 |
| container_title | Diagnostic molecular pathology |
| container_volume | 19 |
| creator | Beaulieu, Martin Desaulniers, Marie Bertrand, Nicolas Deschesnes, Réna G Beaudry, Guillaume Garon, Geneviève Haince, Jean-François Houde, Michel Holzer, Timothy J |
| description | Up to 30% of patients with stage II (pN0) colon cancer develop recurrences, suggesting that the presence of lymph node (LN) metastases escaped detection at histopathologic staging. A simple way to overcome this limitation and to improve staging accuracy is to use reverse transcription-polymerase chain reaction (RT-PCR) to examine a larger fraction or an entire specimen. The Guanylyl cyclase C (GCC) gene is uniquely expressed in apical cells of the gastrointestinal tract. Its expression in colon cancer cells and metastases is conserved. Therefore, detection of GCC mRNA in LNs has been shown to be indicative of the presence of colon cancer metastases. As the current processing of LNs involves formalin fixation and paraffin embedding, we developed a method for extracting RNA from formalin-fixed paraffin-embedded LN specimens and detecting GCC mRNA by quantitative RT-PCR. The assay has a dynamic range of 5 logs, an average amplification efficiency of 98.4% (95% confidence interval, 96.6-100.3), a reaction linearity of 0.998 (95% confidence interval, 0.997-0.999), and also intraplate and interplate CVs of |
| doi_str_mv | 10.1097/PDM.0b013e3181ad5ac3 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_867750108</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>733314127</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3834-88adec0151fd644d617a9e8c3ac217acbf66ebbb0c02dd36f8871591687efec3</originalsourceid><addsrcrecordid>eNqFkU1v1DAQhiMEoqXwDxDyjVPKOI4d57hKu6XSAlG1EkfLcSbagBNvbUcl_x6vWorEhcNoZqT34_Bk2XsKlxTq6lN79eUSOqAMGZVU91wb9iI7p5yxvAQhXqYbeJHXnNOz7E0IPwCAlZK_zs4KoFIAyPPs12bWdo2j0Za06AfnJz0bJG4gmtzf7fO2uSObEPRKoiNXGNFEcrPoebWrJc1qrA5IGjLOZLttr8lunY4H8tX1GE4ZrY4jzjGQ72M8kMZZN5PmVODfZq8GbQO-e9oX2X57vW8-57tvN7fNZpcbJlmZS6l7NEA5HXpRlr2gla5RGqZNkU7TDUJg13VgoOh7JgYpK8prKmSFAxp2kX18jD16d79giGoag0Fr9YxuCUqKquJAQf5XWTHGaEmLKinLR6XxLgSPgzr6cdJ-VRTUiY1KbNS_bJLtw1PB0k3YP5v-wPib--BsRB9-2uUBvTqgtvGgEj1aCxB50ieQ6c3TQMl-A_5mmmk</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>733314127</pqid></control><display><type>article</type><title>Analytical Performance of a qRT-PCR Assay to Detect Guanylyl Cyclase C in FFPE Lymph Nodes of Patients With Colon Cancer</title><source>MEDLINE</source><source>Journals@Ovid Complete</source><creator>Beaulieu, Martin ; Desaulniers, Marie ; Bertrand, Nicolas ; Deschesnes, Réna G ; Beaudry, Guillaume ; Garon, Geneviève ; Haince, Jean-François ; Houde, Michel ; Holzer, Timothy J</creator><creatorcontrib>Beaulieu, Martin ; Desaulniers, Marie ; Bertrand, Nicolas ; Deschesnes, Réna G ; Beaudry, Guillaume ; Garon, Geneviève ; Haince, Jean-François ; Houde, Michel ; Holzer, Timothy J</creatorcontrib><description>Up to 30% of patients with stage II (pN0) colon cancer develop recurrences, suggesting that the presence of lymph node (LN) metastases escaped detection at histopathologic staging. A simple way to overcome this limitation and to improve staging accuracy is to use reverse transcription-polymerase chain reaction (RT-PCR) to examine a larger fraction or an entire specimen. The Guanylyl cyclase C (GCC) gene is uniquely expressed in apical cells of the gastrointestinal tract. Its expression in colon cancer cells and metastases is conserved. Therefore, detection of GCC mRNA in LNs has been shown to be indicative of the presence of colon cancer metastases. As the current processing of LNs involves formalin fixation and paraffin embedding, we developed a method for extracting RNA from formalin-fixed paraffin-embedded LN specimens and detecting GCC mRNA by quantitative RT-PCR. The assay has a dynamic range of 5 logs, an average amplification efficiency of 98.4% (95% confidence interval, 96.6-100.3), a reaction linearity of 0.998 (95% confidence interval, 0.997-0.999), and also intraplate and interplate CVs of <1% and <5%, respectively. The test specificity was 98% with LNs collected from patients affected by conditions other than colon cancer (n=380). Sensitivity was 97% for patients with stage III colon cancer (n=34), whereas 35% of patients with stages I and II disease (n=51) had at least 1 GCC mRNA-positive LN. The high specificity of GCC mRNA suggests that routine utilization of the quantitative RT-PCR test has the potential to improve the detection of colon cancer metastases in LNs.</description><identifier>ISSN: 1052-9551</identifier><identifier>EISSN: 1533-4066</identifier><identifier>DOI: 10.1097/PDM.0b013e3181ad5ac3</identifier><identifier>PMID: 20186008</identifier><language>eng</language><publisher>United States: Lippincott Williams & Wilkins, Inc</publisher><subject>Colonic Neoplasms - diagnosis ; Colonic Neoplasms - secondary ; Fixatives - pharmacology ; Formaldehyde - pharmacology ; Guanylate Cyclase - genetics ; Humans ; Lymph Nodes - pathology ; Paraffin Embedding ; Pathology, Molecular - methods ; Receptors, Enterotoxin ; Receptors, Guanylate Cyclase-Coupled ; Receptors, Peptide - genetics ; Reverse Transcriptase Polymerase Chain Reaction - methods ; Sensitivity and Specificity ; Tissue Fixation</subject><ispartof>Diagnostic molecular pathology, 2010-03, Vol.19 (1), p.20-27</ispartof><rights>2010 Lippincott Williams & Wilkins, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3834-88adec0151fd644d617a9e8c3ac217acbf66ebbb0c02dd36f8871591687efec3</citedby><cites>FETCH-LOGICAL-c3834-88adec0151fd644d617a9e8c3ac217acbf66ebbb0c02dd36f8871591687efec3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20186008$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Beaulieu, Martin</creatorcontrib><creatorcontrib>Desaulniers, Marie</creatorcontrib><creatorcontrib>Bertrand, Nicolas</creatorcontrib><creatorcontrib>Deschesnes, Réna G</creatorcontrib><creatorcontrib>Beaudry, Guillaume</creatorcontrib><creatorcontrib>Garon, Geneviève</creatorcontrib><creatorcontrib>Haince, Jean-François</creatorcontrib><creatorcontrib>Houde, Michel</creatorcontrib><creatorcontrib>Holzer, Timothy J</creatorcontrib><title>Analytical Performance of a qRT-PCR Assay to Detect Guanylyl Cyclase C in FFPE Lymph Nodes of Patients With Colon Cancer</title><title>Diagnostic molecular pathology</title><addtitle>Diagn Mol Pathol</addtitle><description>Up to 30% of patients with stage II (pN0) colon cancer develop recurrences, suggesting that the presence of lymph node (LN) metastases escaped detection at histopathologic staging. A simple way to overcome this limitation and to improve staging accuracy is to use reverse transcription-polymerase chain reaction (RT-PCR) to examine a larger fraction or an entire specimen. The Guanylyl cyclase C (GCC) gene is uniquely expressed in apical cells of the gastrointestinal tract. Its expression in colon cancer cells and metastases is conserved. Therefore, detection of GCC mRNA in LNs has been shown to be indicative of the presence of colon cancer metastases. As the current processing of LNs involves formalin fixation and paraffin embedding, we developed a method for extracting RNA from formalin-fixed paraffin-embedded LN specimens and detecting GCC mRNA by quantitative RT-PCR. The assay has a dynamic range of 5 logs, an average amplification efficiency of 98.4% (95% confidence interval, 96.6-100.3), a reaction linearity of 0.998 (95% confidence interval, 0.997-0.999), and also intraplate and interplate CVs of <1% and <5%, respectively. The test specificity was 98% with LNs collected from patients affected by conditions other than colon cancer (n=380). Sensitivity was 97% for patients with stage III colon cancer (n=34), whereas 35% of patients with stages I and II disease (n=51) had at least 1 GCC mRNA-positive LN. The high specificity of GCC mRNA suggests that routine utilization of the quantitative RT-PCR test has the potential to improve the detection of colon cancer metastases in LNs.</description><subject>Colonic Neoplasms - diagnosis</subject><subject>Colonic Neoplasms - secondary</subject><subject>Fixatives - pharmacology</subject><subject>Formaldehyde - pharmacology</subject><subject>Guanylate Cyclase - genetics</subject><subject>Humans</subject><subject>Lymph Nodes - pathology</subject><subject>Paraffin Embedding</subject><subject>Pathology, Molecular - methods</subject><subject>Receptors, Enterotoxin</subject><subject>Receptors, Guanylate Cyclase-Coupled</subject><subject>Receptors, Peptide - genetics</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>Sensitivity and Specificity</subject><subject>Tissue Fixation</subject><issn>1052-9551</issn><issn>1533-4066</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAQhiMEoqXwDxDyjVPKOI4d57hKu6XSAlG1EkfLcSbagBNvbUcl_x6vWorEhcNoZqT34_Bk2XsKlxTq6lN79eUSOqAMGZVU91wb9iI7p5yxvAQhXqYbeJHXnNOz7E0IPwCAlZK_zs4KoFIAyPPs12bWdo2j0Za06AfnJz0bJG4gmtzf7fO2uSObEPRKoiNXGNFEcrPoebWrJc1qrA5IGjLOZLttr8lunY4H8tX1GE4ZrY4jzjGQ72M8kMZZN5PmVODfZq8GbQO-e9oX2X57vW8-57tvN7fNZpcbJlmZS6l7NEA5HXpRlr2gla5RGqZNkU7TDUJg13VgoOh7JgYpK8prKmSFAxp2kX18jD16d79giGoag0Fr9YxuCUqKquJAQf5XWTHGaEmLKinLR6XxLgSPgzr6cdJ-VRTUiY1KbNS_bJLtw1PB0k3YP5v-wPib--BsRB9-2uUBvTqgtvGgEj1aCxB50ieQ6c3TQMl-A_5mmmk</recordid><startdate>201003</startdate><enddate>201003</enddate><creator>Beaulieu, Martin</creator><creator>Desaulniers, Marie</creator><creator>Bertrand, Nicolas</creator><creator>Deschesnes, Réna G</creator><creator>Beaudry, Guillaume</creator><creator>Garon, Geneviève</creator><creator>Haince, Jean-François</creator><creator>Houde, Michel</creator><creator>Holzer, Timothy J</creator><general>Lippincott Williams & Wilkins, Inc</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>7X8</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>201003</creationdate><title>Analytical Performance of a qRT-PCR Assay to Detect Guanylyl Cyclase C in FFPE Lymph Nodes of Patients With Colon Cancer</title><author>Beaulieu, Martin ; Desaulniers, Marie ; Bertrand, Nicolas ; Deschesnes, Réna G ; Beaudry, Guillaume ; Garon, Geneviève ; Haince, Jean-François ; Houde, Michel ; Holzer, Timothy J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3834-88adec0151fd644d617a9e8c3ac217acbf66ebbb0c02dd36f8871591687efec3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Colonic Neoplasms - diagnosis</topic><topic>Colonic Neoplasms - secondary</topic><topic>Fixatives - pharmacology</topic><topic>Formaldehyde - pharmacology</topic><topic>Guanylate Cyclase - genetics</topic><topic>Humans</topic><topic>Lymph Nodes - pathology</topic><topic>Paraffin Embedding</topic><topic>Pathology, Molecular - methods</topic><topic>Receptors, Enterotoxin</topic><topic>Receptors, Guanylate Cyclase-Coupled</topic><topic>Receptors, Peptide - genetics</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>Sensitivity and Specificity</topic><topic>Tissue Fixation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Beaulieu, Martin</creatorcontrib><creatorcontrib>Desaulniers, Marie</creatorcontrib><creatorcontrib>Bertrand, Nicolas</creatorcontrib><creatorcontrib>Deschesnes, Réna G</creatorcontrib><creatorcontrib>Beaudry, Guillaume</creatorcontrib><creatorcontrib>Garon, Geneviève</creatorcontrib><creatorcontrib>Haince, Jean-François</creatorcontrib><creatorcontrib>Houde, Michel</creatorcontrib><creatorcontrib>Holzer, Timothy J</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Diagnostic molecular pathology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Beaulieu, Martin</au><au>Desaulniers, Marie</au><au>Bertrand, Nicolas</au><au>Deschesnes, Réna G</au><au>Beaudry, Guillaume</au><au>Garon, Geneviève</au><au>Haince, Jean-François</au><au>Houde, Michel</au><au>Holzer, Timothy J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Analytical Performance of a qRT-PCR Assay to Detect Guanylyl Cyclase C in FFPE Lymph Nodes of Patients With Colon Cancer</atitle><jtitle>Diagnostic molecular pathology</jtitle><addtitle>Diagn Mol Pathol</addtitle><date>2010-03</date><risdate>2010</risdate><volume>19</volume><issue>1</issue><spage>20</spage><epage>27</epage><pages>20-27</pages><issn>1052-9551</issn><eissn>1533-4066</eissn><abstract>Up to 30% of patients with stage II (pN0) colon cancer develop recurrences, suggesting that the presence of lymph node (LN) metastases escaped detection at histopathologic staging. A simple way to overcome this limitation and to improve staging accuracy is to use reverse transcription-polymerase chain reaction (RT-PCR) to examine a larger fraction or an entire specimen. The Guanylyl cyclase C (GCC) gene is uniquely expressed in apical cells of the gastrointestinal tract. Its expression in colon cancer cells and metastases is conserved. Therefore, detection of GCC mRNA in LNs has been shown to be indicative of the presence of colon cancer metastases. As the current processing of LNs involves formalin fixation and paraffin embedding, we developed a method for extracting RNA from formalin-fixed paraffin-embedded LN specimens and detecting GCC mRNA by quantitative RT-PCR. The assay has a dynamic range of 5 logs, an average amplification efficiency of 98.4% (95% confidence interval, 96.6-100.3), a reaction linearity of 0.998 (95% confidence interval, 0.997-0.999), and also intraplate and interplate CVs of <1% and <5%, respectively. The test specificity was 98% with LNs collected from patients affected by conditions other than colon cancer (n=380). Sensitivity was 97% for patients with stage III colon cancer (n=34), whereas 35% of patients with stages I and II disease (n=51) had at least 1 GCC mRNA-positive LN. The high specificity of GCC mRNA suggests that routine utilization of the quantitative RT-PCR test has the potential to improve the detection of colon cancer metastases in LNs.</abstract><cop>United States</cop><pub>Lippincott Williams & Wilkins, Inc</pub><pmid>20186008</pmid><doi>10.1097/PDM.0b013e3181ad5ac3</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1052-9551 |
| ispartof | Diagnostic molecular pathology, 2010-03, Vol.19 (1), p.20-27 |
| issn | 1052-9551 1533-4066 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_867750108 |
| source | MEDLINE; Journals@Ovid Complete |
| subjects | Colonic Neoplasms - diagnosis Colonic Neoplasms - secondary Fixatives - pharmacology Formaldehyde - pharmacology Guanylate Cyclase - genetics Humans Lymph Nodes - pathology Paraffin Embedding Pathology, Molecular - methods Receptors, Enterotoxin Receptors, Guanylate Cyclase-Coupled Receptors, Peptide - genetics Reverse Transcriptase Polymerase Chain Reaction - methods Sensitivity and Specificity Tissue Fixation |
| title | Analytical Performance of a qRT-PCR Assay to Detect Guanylyl Cyclase C in FFPE Lymph Nodes of Patients With Colon Cancer |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-13T07%3A36%3A47IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Analytical%20Performance%20of%20a%20qRT-PCR%20Assay%20to%20Detect%20Guanylyl%20Cyclase%20C%20in%20FFPE%20Lymph%20Nodes%20of%20Patients%20With%20Colon%20Cancer&rft.jtitle=Diagnostic%20molecular%20pathology&rft.au=Beaulieu,%20Martin&rft.date=2010-03&rft.volume=19&rft.issue=1&rft.spage=20&rft.epage=27&rft.pages=20-27&rft.issn=1052-9551&rft.eissn=1533-4066&rft_id=info:doi/10.1097/PDM.0b013e3181ad5ac3&rft_dat=%3Cproquest_cross%3E733314127%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=733314127&rft_id=info:pmid/20186008&rfr_iscdi=true |