Real-time PCR method for the quantitative analysis of human T-cell receptor γ and β gene rearrangements
Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the αβ vs. γδ lineage decision, and characterizing T-cell leukemias. Methods such as PCR and quantitative Southern blotting provide useful in...
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| Veröffentlicht in: | Journal of immunological methods 2005-05, Vol.300 (1), p.12-23 |
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| container_title | Journal of immunological methods |
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| creator | Chain, J.L. Joachims, M.L. Hooker, S.W. Laurent, A.B. Knott-Craig, C.K. Thompson, L.F. |
| description | Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the αβ vs. γδ lineage decision, and characterizing T-cell leukemias. Methods such as PCR and quantitative Southern blotting provide useful information, but also have significant shortcomings such as lack of quantitation in the case of PCR and technical challenges in the case of Southern blotting. Here we describe a real-time PCR method that overcomes many of these shortcomings. This new method shows comparable results for the fraction of unrearranged TCRγ and TCRβ genes in human thymocytes and peripheral blood T cells as Southern blotting, and has the advantages of being simple to perform, highly quantitative, and requiring nanogram quantities of DNA. We also describe a real-time PCR method to quantitate T-cell receptor excision circles formed during TCRβ rearrangements. |
| doi_str_mv | 10.1016/j.jim.2005.01.016 |
| format | Article |
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Methods such as PCR and quantitative Southern blotting provide useful information, but also have significant shortcomings such as lack of quantitation in the case of PCR and technical challenges in the case of Southern blotting. Here we describe a real-time PCR method that overcomes many of these shortcomings. This new method shows comparable results for the fraction of unrearranged TCRγ and TCRβ genes in human thymocytes and peripheral blood T cells as Southern blotting, and has the advantages of being simple to perform, highly quantitative, and requiring nanogram quantities of DNA. We also describe a real-time PCR method to quantitate T-cell receptor excision circles formed during TCRβ rearrangements.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/j.jim.2005.01.016</identifier><identifier>PMID: 15882867</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Base Sequence ; Biological and medical sciences ; Blotting, Southern ; Child ; DNA - analysis ; DNA - genetics ; DNA Primers - genetics ; DNA Probes - genetics ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Infant ; Molecular immunology ; Molecular Probe Techniques ; Polymerase Chain Reaction - methods ; Quantitative Southern blot ; Real-time PCR ; T-cell receptor excision circle ; T-cell receptor gene rearrangement ; T-Lymphocytes - immunology ; Techniques</subject><ispartof>Journal of immunological methods, 2005-05, Vol.300 (1), p.12-23</ispartof><rights>2005 Elsevier B.V.</rights><rights>2005 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c541t-3455a1c16cee2ffd9b6ddda7d6a06c8603f271be92ae7fe7e0401d24f5d81cc83</citedby><cites>FETCH-LOGICAL-c541t-3455a1c16cee2ffd9b6ddda7d6a06c8603f271be92ae7fe7e0401d24f5d81cc83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jim.2005.01.016$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=16884265$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15882867$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chain, J.L.</creatorcontrib><creatorcontrib>Joachims, M.L.</creatorcontrib><creatorcontrib>Hooker, S.W.</creatorcontrib><creatorcontrib>Laurent, A.B.</creatorcontrib><creatorcontrib>Knott-Craig, C.K.</creatorcontrib><creatorcontrib>Thompson, L.F.</creatorcontrib><title>Real-time PCR method for the quantitative analysis of human T-cell receptor γ and β gene rearrangements</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the αβ vs. γδ lineage decision, and characterizing T-cell leukemias. Methods such as PCR and quantitative Southern blotting provide useful information, but also have significant shortcomings such as lack of quantitation in the case of PCR and technical challenges in the case of Southern blotting. Here we describe a real-time PCR method that overcomes many of these shortcomings. This new method shows comparable results for the fraction of unrearranged TCRγ and TCRβ genes in human thymocytes and peripheral blood T cells as Southern blotting, and has the advantages of being simple to perform, highly quantitative, and requiring nanogram quantities of DNA. We also describe a real-time PCR method to quantitate T-cell receptor excision circles formed during TCRβ rearrangements.</description><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Southern</subject><subject>Child</subject><subject>DNA - analysis</subject><subject>DNA - genetics</subject><subject>DNA Primers - genetics</subject><subject>DNA Probes - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Gene Rearrangement, beta-Chain T-Cell Antigen Receptor</subject><subject>Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor</subject><subject>Humans</subject><subject>Infant</subject><subject>Molecular immunology</subject><subject>Molecular Probe Techniques</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Quantitative Southern blot</subject><subject>Real-time PCR</subject><subject>T-cell receptor excision circle</subject><subject>T-cell receptor gene rearrangement</subject><subject>T-Lymphocytes - immunology</subject><subject>Techniques</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkd2KEzEUgIMobl19AG8kN3o3NSedJDMIgpT1BxaUZb0OaXLSpsxPN8kU9rX0PfaZNqXF1RsXDgSS7_zlI-Q1sDkwkO-3823o55wxMWdQQj4hM2gUr1TLxFMyY4zzCpRoz8iLlLaMMWCSPSdnIJqGN1LNSLhC01U59Eh_LK9oj3kzOurHSPMG6c1khhyyyWGP1Aymu00h0dHTzdSbgV5XFruORrS4yyXl7neBHL37Rdc4YLk3MZphjT0OOb0kz7zpEr46nefk5-eL6-XX6vL7l2_LT5eVFTXkalELYcCCtIjce9eupHPOKCcNk7aRbOG5ghW23KDyqJDVDByvvXANWNsszsnHY93dtOrR2dI7mk7vYuhNvNWjCfrflyFs9Hrcazh8SbsoBd6dCsTxZsKUdR_SYVEz4DglLVWBangchFbwlsv6cVAJAXzBCghH0MYxpYj-z9jA9EG53uqiXB-UawYlZMl58_e-DxknxwV4ewJMsqbzRYkN6YGTTVNzKQr34chhsbMPGHWyAQeLLhTDWbsx_GeMe5_HzJM</recordid><startdate>20050501</startdate><enddate>20050501</enddate><creator>Chain, J.L.</creator><creator>Joachims, M.L.</creator><creator>Hooker, S.W.</creator><creator>Laurent, A.B.</creator><creator>Knott-Craig, C.K.</creator><creator>Thompson, L.F.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</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>7T5</scope><scope>H94</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20050501</creationdate><title>Real-time PCR method for the quantitative analysis of human T-cell receptor γ and β gene rearrangements</title><author>Chain, J.L. ; Joachims, M.L. ; Hooker, S.W. ; Laurent, A.B. ; Knott-Craig, C.K. ; Thompson, L.F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c541t-3455a1c16cee2ffd9b6ddda7d6a06c8603f271be92ae7fe7e0401d24f5d81cc83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Southern</topic><topic>Child</topic><topic>DNA - analysis</topic><topic>DNA - genetics</topic><topic>DNA Primers - genetics</topic><topic>DNA Probes - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Gene Rearrangement, beta-Chain T-Cell Antigen Receptor</topic><topic>Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor</topic><topic>Humans</topic><topic>Infant</topic><topic>Molecular immunology</topic><topic>Molecular Probe Techniques</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Quantitative Southern blot</topic><topic>Real-time PCR</topic><topic>T-cell receptor excision circle</topic><topic>T-cell receptor gene rearrangement</topic><topic>T-Lymphocytes - immunology</topic><topic>Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chain, J.L.</creatorcontrib><creatorcontrib>Joachims, M.L.</creatorcontrib><creatorcontrib>Hooker, S.W.</creatorcontrib><creatorcontrib>Laurent, A.B.</creatorcontrib><creatorcontrib>Knott-Craig, C.K.</creatorcontrib><creatorcontrib>Thompson, L.F.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chain, J.L.</au><au>Joachims, M.L.</au><au>Hooker, S.W.</au><au>Laurent, A.B.</au><au>Knott-Craig, C.K.</au><au>Thompson, L.F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Real-time PCR method for the quantitative analysis of human T-cell receptor γ and β gene rearrangements</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>2005-05-01</date><risdate>2005</risdate><volume>300</volume><issue>1</issue><spage>12</spage><epage>23</epage><pages>12-23</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the αβ vs. γδ lineage decision, and characterizing T-cell leukemias. 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| ispartof | Journal of immunological methods, 2005-05, Vol.300 (1), p.12-23 |
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| subjects | Base Sequence Biological and medical sciences Blotting, Southern Child DNA - analysis DNA - genetics DNA Primers - genetics DNA Probes - genetics Fundamental and applied biological sciences. Psychology Fundamental immunology Gene Rearrangement, beta-Chain T-Cell Antigen Receptor Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor Humans Infant Molecular immunology Molecular Probe Techniques Polymerase Chain Reaction - methods Quantitative Southern blot Real-time PCR T-cell receptor excision circle T-cell receptor gene rearrangement T-Lymphocytes - immunology Techniques |
| title | Real-time PCR method for the quantitative analysis of human T-cell receptor γ and β gene rearrangements |
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