Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay
A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood mononuclear cells (PBMCs). The HTLV-I copy number was referred to the actual amount of cellular DNA by means of the quantitation of the albumin gene. Ten co...
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| Veröffentlicht in: | Journal of virological methods 2002-04, Vol.102 (1), p.37-51 |
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| creator | Dehée, Axelle Césaire, Raymond Désiré, Nathalie Lézin, Agnès Bourdonné, Olivier Béra, Odile Plumelle, Yves Smadja, Didier Nicolas, Jean-Claude |
| description | A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood mononuclear cells (PBMCs). The HTLV-I copy number was referred to the actual amount of cellular DNA by means of the quantitation of the albumin gene. Ten copies of HTLV-I DNA could be detected with 100% sensitivity, and the assay had a wide range of at least 5
log
10. Intra- and inter-assay reproducibility was evaluated using independent extractions of PBMCs from an HTLV-I-infected patient (coefficients of variation, 24 and 7% respectively). The performance of this TaqMan PCR assay, coupled with its high throughput, thus allows reliable routine follow-up of HTLV-I proviral load in infected patients. Preliminary results using clinical samples indicate a higher proviral load in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis than in asymptomatic carriers, and also suggest the usefulness of this quantitative measurement to assess the etiological link between HTLV-I and adult T-cell leukaemia/lymphoma-like syndromes. |
| doi_str_mv | 10.1016/S0166-0934(01)00445-1 |
| format | Article |
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log
10. Intra- and inter-assay reproducibility was evaluated using independent extractions of PBMCs from an HTLV-I-infected patient (coefficients of variation, 24 and 7% respectively). The performance of this TaqMan PCR assay, coupled with its high throughput, thus allows reliable routine follow-up of HTLV-I proviral load in infected patients. Preliminary results using clinical samples indicate a higher proviral load in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis than in asymptomatic carriers, and also suggest the usefulness of this quantitative measurement to assess the etiological link between HTLV-I and adult T-cell leukaemia/lymphoma-like syndromes.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/S0166-0934(01)00445-1</identifier><identifier>PMID: 11879691</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Base Sequence ; Biological and medical sciences ; Carrier State ; DNA, Viral ; Fundamental and applied biological sciences. Psychology ; HTLV-I ; Human T-lymphotropic virus 1 ; Human T-lymphotropic virus 1 - genetics ; Human T-lymphotropic virus 1 - growth & development ; Human viral diseases ; Humans ; Infectious diseases ; Leukemia-Lymphoma, Adult T-Cell - blood ; Leukemia-Lymphoma, Adult T-Cell - virology ; Leukocytes, Mononuclear - virology ; Medical sciences ; Microbiology ; Molecular Sequence Data ; Paraparesis, Tropical Spastic - blood ; Paraparesis, Tropical Spastic - virology ; Polymerase Chain Reaction - methods ; Proviruses - genetics ; Proviruses - growth & development ; Real-time quantitative PCR ; Reproducibility of Results ; Sensitivity and Specificity ; Taq Polymerase ; TaqMan ; Techniques used in virology ; Viral diseases ; Viral diseases of the lymphoid tissue and the blood. Aids ; Viral Load ; Virology</subject><ispartof>Journal of virological methods, 2002-04, Vol.102 (1), p.37-51</ispartof><rights>2002 Elsevier Science B.V.</rights><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-f0534d00dc5294a4bd460f6a64b1932d2a03df6632f6132421b7e2e6fa737c8e3</citedby><cites>FETCH-LOGICAL-c474t-f0534d00dc5294a4bd460f6a64b1932d2a03df6632f6132421b7e2e6fa737c8e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0166093401004451$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=13535240$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11879691$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Dehée, Axelle</creatorcontrib><creatorcontrib>Césaire, Raymond</creatorcontrib><creatorcontrib>Désiré, Nathalie</creatorcontrib><creatorcontrib>Lézin, Agnès</creatorcontrib><creatorcontrib>Bourdonné, Olivier</creatorcontrib><creatorcontrib>Béra, Odile</creatorcontrib><creatorcontrib>Plumelle, Yves</creatorcontrib><creatorcontrib>Smadja, Didier</creatorcontrib><creatorcontrib>Nicolas, Jean-Claude</creatorcontrib><title>Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood mononuclear cells (PBMCs). The HTLV-I copy number was referred to the actual amount of cellular DNA by means of the quantitation of the albumin gene. Ten copies of HTLV-I DNA could be detected with 100% sensitivity, and the assay had a wide range of at least 5
log
10. Intra- and inter-assay reproducibility was evaluated using independent extractions of PBMCs from an HTLV-I-infected patient (coefficients of variation, 24 and 7% respectively). The performance of this TaqMan PCR assay, coupled with its high throughput, thus allows reliable routine follow-up of HTLV-I proviral load in infected patients. Preliminary results using clinical samples indicate a higher proviral load in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis than in asymptomatic carriers, and also suggest the usefulness of this quantitative measurement to assess the etiological link between HTLV-I and adult T-cell leukaemia/lymphoma-like syndromes.</description><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Carrier State</subject><subject>DNA, Viral</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HTLV-I</subject><subject>Human T-lymphotropic virus 1</subject><subject>Human T-lymphotropic virus 1 - genetics</subject><subject>Human T-lymphotropic virus 1 - growth & development</subject><subject>Human viral diseases</subject><subject>Humans</subject><subject>Infectious diseases</subject><subject>Leukemia-Lymphoma, Adult T-Cell - blood</subject><subject>Leukemia-Lymphoma, Adult T-Cell - virology</subject><subject>Leukocytes, Mononuclear - virology</subject><subject>Medical sciences</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Paraparesis, Tropical Spastic - blood</subject><subject>Paraparesis, Tropical Spastic - virology</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Proviruses - genetics</subject><subject>Proviruses - growth & development</subject><subject>Real-time quantitative PCR</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Taq Polymerase</subject><subject>TaqMan</subject><subject>Techniques used in virology</subject><subject>Viral diseases</subject><subject>Viral diseases of the lymphoid tissue and the blood. Aids</subject><subject>Viral Load</subject><subject>Virology</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1r20AQhpeQ0jhpf0LDXhLSg9LZD62kUylOUhsc0g-n12W0H7BBluxdOeB_Hzk29dGXGRiemXl5CPnC4JYBU9_-DkVlUAl5A-wrgJR5xk7IiJVFNYxLeUpG_5Ezcp7SCwDkhRAfyRnbUqpiI3L3e41tH3rsQ9fSztPJfPYvm9Jl7F5DxIY2HVpabyjSOa4esaXRYZP1YeHor_Efiinh5hP54LFJ7vO-X5Dnh_v5eJLNnn5Oxz9mmZGF7DMPuZAWwJqcVxJlbaUCr1DJmlWCW44grFdKcK-Y4JKzunDcKY-FKEzpxAW53t0d0q3WLvV6EZJxTYOt69ZJF0xWVQVwFGQlL0FxNYD5DjSxSyk6r5cxLDBuNAO99azfPeutRA1Mv3vWbNi73D9Y1wtnD1t7sQNwtQcwGWx8xNaEdOBELnIut0m_7zg3eHsNLupkgmuNsyE602vbhSNR3gBIYZa4</recordid><startdate>20020401</startdate><enddate>20020401</enddate><creator>Dehée, Axelle</creator><creator>Césaire, Raymond</creator><creator>Désiré, Nathalie</creator><creator>Lézin, Agnès</creator><creator>Bourdonné, Olivier</creator><creator>Béra, Odile</creator><creator>Plumelle, Yves</creator><creator>Smadja, Didier</creator><creator>Nicolas, Jean-Claude</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>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20020401</creationdate><title>Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay</title><author>Dehée, Axelle ; Césaire, Raymond ; Désiré, Nathalie ; Lézin, Agnès ; Bourdonné, Olivier ; Béra, Odile ; Plumelle, Yves ; Smadja, Didier ; Nicolas, Jean-Claude</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-f0534d00dc5294a4bd460f6a64b1932d2a03df6632f6132421b7e2e6fa737c8e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Carrier State</topic><topic>DNA, Viral</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HTLV-I</topic><topic>Human T-lymphotropic virus 1</topic><topic>Human T-lymphotropic virus 1 - genetics</topic><topic>Human T-lymphotropic virus 1 - growth & development</topic><topic>Human viral diseases</topic><topic>Humans</topic><topic>Infectious diseases</topic><topic>Leukemia-Lymphoma, Adult T-Cell - blood</topic><topic>Leukemia-Lymphoma, Adult T-Cell - virology</topic><topic>Leukocytes, Mononuclear - virology</topic><topic>Medical sciences</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Paraparesis, Tropical Spastic - blood</topic><topic>Paraparesis, Tropical Spastic - virology</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Proviruses - genetics</topic><topic>Proviruses - growth & development</topic><topic>Real-time quantitative PCR</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Taq Polymerase</topic><topic>TaqMan</topic><topic>Techniques used in virology</topic><topic>Viral diseases</topic><topic>Viral diseases of the lymphoid tissue and the blood. Aids</topic><topic>Viral Load</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dehée, Axelle</creatorcontrib><creatorcontrib>Césaire, Raymond</creatorcontrib><creatorcontrib>Désiré, Nathalie</creatorcontrib><creatorcontrib>Lézin, Agnès</creatorcontrib><creatorcontrib>Bourdonné, Olivier</creatorcontrib><creatorcontrib>Béra, Odile</creatorcontrib><creatorcontrib>Plumelle, Yves</creatorcontrib><creatorcontrib>Smadja, Didier</creatorcontrib><creatorcontrib>Nicolas, Jean-Claude</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>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dehée, Axelle</au><au>Césaire, Raymond</au><au>Désiré, Nathalie</au><au>Lézin, Agnès</au><au>Bourdonné, Olivier</au><au>Béra, Odile</au><au>Plumelle, Yves</au><au>Smadja, Didier</au><au>Nicolas, Jean-Claude</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2002-04-01</date><risdate>2002</risdate><volume>102</volume><issue>1</issue><spage>37</spage><epage>51</epage><pages>37-51</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood mononuclear cells (PBMCs). The HTLV-I copy number was referred to the actual amount of cellular DNA by means of the quantitation of the albumin gene. Ten copies of HTLV-I DNA could be detected with 100% sensitivity, and the assay had a wide range of at least 5
log
10. Intra- and inter-assay reproducibility was evaluated using independent extractions of PBMCs from an HTLV-I-infected patient (coefficients of variation, 24 and 7% respectively). The performance of this TaqMan PCR assay, coupled with its high throughput, thus allows reliable routine follow-up of HTLV-I proviral load in infected patients. Preliminary results using clinical samples indicate a higher proviral load in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis than in asymptomatic carriers, and also suggest the usefulness of this quantitative measurement to assess the etiological link between HTLV-I and adult T-cell leukaemia/lymphoma-like syndromes.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>11879691</pmid><doi>10.1016/S0166-0934(01)00445-1</doi><tpages>15</tpages></addata></record> |
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| subjects | Base Sequence Biological and medical sciences Carrier State DNA, Viral Fundamental and applied biological sciences. Psychology HTLV-I Human T-lymphotropic virus 1 Human T-lymphotropic virus 1 - genetics Human T-lymphotropic virus 1 - growth & development Human viral diseases Humans Infectious diseases Leukemia-Lymphoma, Adult T-Cell - blood Leukemia-Lymphoma, Adult T-Cell - virology Leukocytes, Mononuclear - virology Medical sciences Microbiology Molecular Sequence Data Paraparesis, Tropical Spastic - blood Paraparesis, Tropical Spastic - virology Polymerase Chain Reaction - methods Proviruses - genetics Proviruses - growth & development Real-time quantitative PCR Reproducibility of Results Sensitivity and Specificity Taq Polymerase TaqMan Techniques used in virology Viral diseases Viral diseases of the lymphoid tissue and the blood. Aids Viral Load Virology |
| title | Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay |
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