Evaluation of the Roche COBAS® TaqMan® HIV-1 test for quantifying HIV-1 RNA in infected cells and lymphoid tissue
Lymphoid tissue is the main reservoir of HIV-1 in infected individuals. In this study the COBAS® TaqMan® HIV-1 test was evaluated for use with the High Pure System (HPS), for quantifying HIV-1 RNA in infected cells and lymphoid tissue specimens. Serial dilutions of 8E5–LAV1 infected T-cells into SUP...
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| Veröffentlicht in: | Journal of virological methods 2011-06, Vol.174 (1-2), p.69-76 |
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| container_title | Journal of virological methods |
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| creator | Gil, Cristina Garcia, M. Teresa Garcia, Felipe Miró, José M. Agüero, Fernando Alós, Lucía Zamora, Laura Capón, Alicia Costa, Josep Pumarola, Tomás Gatell, José M. |
| description | Lymphoid tissue is the main reservoir of HIV-1 in infected individuals. In this study the COBAS® TaqMan® HIV-1 test was evaluated for use with the High Pure System (HPS), for quantifying HIV-1 RNA in infected cells and lymphoid tissue specimens. Serial dilutions of 8E5–LAV1 infected T-cells into SUP-T1 cells and 44 tonsil specimens were examined. Some modifications of the test were required, such as the removal of residual DNA and the HIV-1 RNA output copies were adjusted to the sample input and expressed as HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan HIV-1® (HPS) test proved to be a robust, sensitive, specific and reproducible method for quantifying HIV-1 RNA in infected cells and lymphoid tissue. Linearity and reproducibility were observed in serial dilutions of 8E5–LAV1 infected T-cells (R2>0.86). High reproducibility was found in clinical tonsil specimens (Wilcoxon test p>0.05). rDNase I treatment was essential to avoid false positives caused by residual HIV-1 DNA, mainly in tonsil specimens obtained from infected patients receiving effective antiretroviral treatment. Probit analysis determined the limit of detection as 22HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan® HIV-1 (HPS) test thereby proved to be a helpful tool for measuring the HIV-1 viral load in infected cells and lymphoid tissue reservoirs. |
| doi_str_mv | 10.1016/j.jviromet.2011.03.026 |
| format | Article |
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Teresa ; Garcia, Felipe ; Miró, José M. ; Agüero, Fernando ; Alós, Lucía ; Zamora, Laura ; Capón, Alicia ; Costa, Josep ; Pumarola, Tomás ; Gatell, José M.</creator><creatorcontrib>Gil, Cristina ; Garcia, M. Teresa ; Garcia, Felipe ; Miró, José M. ; Agüero, Fernando ; Alós, Lucía ; Zamora, Laura ; Capón, Alicia ; Costa, Josep ; Pumarola, Tomás ; Gatell, José M.</creatorcontrib><description>Lymphoid tissue is the main reservoir of HIV-1 in infected individuals. In this study the COBAS® TaqMan® HIV-1 test was evaluated for use with the High Pure System (HPS), for quantifying HIV-1 RNA in infected cells and lymphoid tissue specimens. Serial dilutions of 8E5–LAV1 infected T-cells into SUP-T1 cells and 44 tonsil specimens were examined. Some modifications of the test were required, such as the removal of residual DNA and the HIV-1 RNA output copies were adjusted to the sample input and expressed as HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan HIV-1® (HPS) test proved to be a robust, sensitive, specific and reproducible method for quantifying HIV-1 RNA in infected cells and lymphoid tissue. Linearity and reproducibility were observed in serial dilutions of 8E5–LAV1 infected T-cells (R2>0.86). High reproducibility was found in clinical tonsil specimens (Wilcoxon test p>0.05). rDNase I treatment was essential to avoid false positives caused by residual HIV-1 DNA, mainly in tonsil specimens obtained from infected patients receiving effective antiretroviral treatment. Probit analysis determined the limit of detection as 22HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan® HIV-1 (HPS) test thereby proved to be a helpful tool for measuring the HIV-1 viral load in infected cells and lymphoid tissue reservoirs.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/j.jviromet.2011.03.026</identifier><identifier>PMID: 21458494</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>Kidlington: Elsevier B.V</publisher><subject>antiretroviral agents ; Biological and medical sciences ; Cells, Cultured ; COBAS TaqMan HIV-1 ; detection limit ; DNA ; Fundamental and applied biological sciences. Psychology ; HIV infections ; HIV-1 - genetics ; HIV-1 - isolation & purification ; HIV-1 RNA quantitation ; Human immunodeficiency virus 1 ; Humans ; Lymphoid tissue ; Microbiology ; Nucleic Acid Amplification Techniques - methods ; Palatine Tonsil - virology ; patients ; probit analysis ; Reagent Kits, Diagnostic ; Reproducibility of Results ; RNA ; RNA, Viral - genetics ; RNA, Viral - isolation & purification ; Sensitivity and Specificity ; T-lymphocytes ; T-Lymphocytes - virology ; Techniques used in virology ; tonsils ; viral load ; Viral Load - methods ; Virology</subject><ispartof>Journal of virological methods, 2011-06, Vol.174 (1-2), p.69-76</ispartof><rights>2011 Elsevier B.V.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c421t-f78e22d8a4b95644788c2b27c11c8608873c399f5e464b3c5a60016253daafa03</citedby><cites>FETCH-LOGICAL-c421t-f78e22d8a4b95644788c2b27c11c8608873c399f5e464b3c5a60016253daafa03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jviromet.2011.03.026$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24196574$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21458494$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gil, Cristina</creatorcontrib><creatorcontrib>Garcia, M. Teresa</creatorcontrib><creatorcontrib>Garcia, Felipe</creatorcontrib><creatorcontrib>Miró, José M.</creatorcontrib><creatorcontrib>Agüero, Fernando</creatorcontrib><creatorcontrib>Alós, Lucía</creatorcontrib><creatorcontrib>Zamora, Laura</creatorcontrib><creatorcontrib>Capón, Alicia</creatorcontrib><creatorcontrib>Costa, Josep</creatorcontrib><creatorcontrib>Pumarola, Tomás</creatorcontrib><creatorcontrib>Gatell, José M.</creatorcontrib><title>Evaluation of the Roche COBAS® TaqMan® HIV-1 test for quantifying HIV-1 RNA in infected cells and lymphoid tissue</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>Lymphoid tissue is the main reservoir of HIV-1 in infected individuals. In this study the COBAS® TaqMan® HIV-1 test was evaluated for use with the High Pure System (HPS), for quantifying HIV-1 RNA in infected cells and lymphoid tissue specimens. Serial dilutions of 8E5–LAV1 infected T-cells into SUP-T1 cells and 44 tonsil specimens were examined. Some modifications of the test were required, such as the removal of residual DNA and the HIV-1 RNA output copies were adjusted to the sample input and expressed as HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan HIV-1® (HPS) test proved to be a robust, sensitive, specific and reproducible method for quantifying HIV-1 RNA in infected cells and lymphoid tissue. Linearity and reproducibility were observed in serial dilutions of 8E5–LAV1 infected T-cells (R2>0.86). High reproducibility was found in clinical tonsil specimens (Wilcoxon test p>0.05). rDNase I treatment was essential to avoid false positives caused by residual HIV-1 DNA, mainly in tonsil specimens obtained from infected patients receiving effective antiretroviral treatment. Probit analysis determined the limit of detection as 22HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan® HIV-1 (HPS) test thereby proved to be a helpful tool for measuring the HIV-1 viral load in infected cells and lymphoid tissue reservoirs.</description><subject>antiretroviral agents</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>COBAS TaqMan HIV-1</subject><subject>detection limit</subject><subject>DNA</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HIV infections</subject><subject>HIV-1 - genetics</subject><subject>HIV-1 - isolation & purification</subject><subject>HIV-1 RNA quantitation</subject><subject>Human immunodeficiency virus 1</subject><subject>Humans</subject><subject>Lymphoid tissue</subject><subject>Microbiology</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>Palatine Tonsil - virology</subject><subject>patients</subject><subject>probit analysis</subject><subject>Reagent Kits, Diagnostic</subject><subject>Reproducibility of Results</subject><subject>RNA</subject><subject>RNA, Viral - genetics</subject><subject>RNA, Viral - isolation & purification</subject><subject>Sensitivity and Specificity</subject><subject>T-lymphocytes</subject><subject>T-Lymphocytes - virology</subject><subject>Techniques used in virology</subject><subject>tonsils</subject><subject>viral load</subject><subject>Viral Load - methods</subject><subject>Virology</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkdtu0zAYgC0EYt3gFYZvEFcJPsVx7ijVYJMGk3bg1vrr2JurNG7tpFJfiofYk-GoHVwiWfYv-fsP_ozQOSUlJVR-XpWrnY9hbYeSEUpLwkvC5Cs0o6puCtIo8RrNMihzzMUJOk1pRQipas7fohNGRaVEI2YoXeygG2HwocfB4eHJ4ttg8r64-Tq_e_6N72H7A_ocXF79KigebBqwCxFvR-gH7_a-fzxe3f6cY9_n5awZbIuN7bqEoW9xt19vnoJv8eBTGu079MZBl-z743mGHr5d3C8ui-ub71eL-XVhBKND4WplGWsViGVTSSFqpQxbstpQapQkStXc8KZxlRVSLLmpQJL8YlbxFsAB4Wfo06HuJobtmAfXa5-mqaC3YUxayZoz3lCZSXkgTQwpRev0Jvo1xL2mRE--9Uq_-NaTb024zr5z4vmxxbhc2_Zv2ovgDHw8ApAMdC5Cb3z6xwnayKqeuA8HzkHQ8Bgz83CXO8np0xRvplZfDoTNynbeRp2Mt72xrY9ZuG6D_9-0fwA5v6pi</recordid><startdate>20110601</startdate><enddate>20110601</enddate><creator>Gil, Cristina</creator><creator>Garcia, M. Teresa</creator><creator>Garcia, Felipe</creator><creator>Miró, José M.</creator><creator>Agüero, Fernando</creator><creator>Alós, Lucía</creator><creator>Zamora, Laura</creator><creator>Capón, Alicia</creator><creator>Costa, Josep</creator><creator>Pumarola, Tomás</creator><creator>Gatell, José M.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>FBQ</scope><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>7X8</scope></search><sort><creationdate>20110601</creationdate><title>Evaluation of the Roche COBAS® TaqMan® HIV-1 test for quantifying HIV-1 RNA in infected cells and lymphoid tissue</title><author>Gil, Cristina ; Garcia, M. Teresa ; Garcia, Felipe ; Miró, José M. ; Agüero, Fernando ; Alós, Lucía ; Zamora, Laura ; Capón, Alicia ; Costa, Josep ; Pumarola, Tomás ; Gatell, José M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c421t-f78e22d8a4b95644788c2b27c11c8608873c399f5e464b3c5a60016253daafa03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>antiretroviral agents</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>COBAS TaqMan HIV-1</topic><topic>detection limit</topic><topic>DNA</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HIV infections</topic><topic>HIV-1 - genetics</topic><topic>HIV-1 - isolation & purification</topic><topic>HIV-1 RNA quantitation</topic><topic>Human immunodeficiency virus 1</topic><topic>Humans</topic><topic>Lymphoid tissue</topic><topic>Microbiology</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>Palatine Tonsil - virology</topic><topic>patients</topic><topic>probit analysis</topic><topic>Reagent Kits, Diagnostic</topic><topic>Reproducibility of Results</topic><topic>RNA</topic><topic>RNA, Viral - genetics</topic><topic>RNA, Viral - isolation & purification</topic><topic>Sensitivity and Specificity</topic><topic>T-lymphocytes</topic><topic>T-Lymphocytes - virology</topic><topic>Techniques used in virology</topic><topic>tonsils</topic><topic>viral load</topic><topic>Viral Load - methods</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gil, Cristina</creatorcontrib><creatorcontrib>Garcia, M. Teresa</creatorcontrib><creatorcontrib>Garcia, Felipe</creatorcontrib><creatorcontrib>Miró, José M.</creatorcontrib><creatorcontrib>Agüero, Fernando</creatorcontrib><creatorcontrib>Alós, Lucía</creatorcontrib><creatorcontrib>Zamora, Laura</creatorcontrib><creatorcontrib>Capón, Alicia</creatorcontrib><creatorcontrib>Costa, Josep</creatorcontrib><creatorcontrib>Pumarola, Tomás</creatorcontrib><creatorcontrib>Gatell, José M.</creatorcontrib><collection>AGRIS</collection><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>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gil, Cristina</au><au>Garcia, M. Teresa</au><au>Garcia, Felipe</au><au>Miró, José M.</au><au>Agüero, Fernando</au><au>Alós, Lucía</au><au>Zamora, Laura</au><au>Capón, Alicia</au><au>Costa, Josep</au><au>Pumarola, Tomás</au><au>Gatell, José M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evaluation of the Roche COBAS® TaqMan® HIV-1 test for quantifying HIV-1 RNA in infected cells and lymphoid tissue</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2011-06-01</date><risdate>2011</risdate><volume>174</volume><issue>1-2</issue><spage>69</spage><epage>76</epage><pages>69-76</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>Lymphoid tissue is the main reservoir of HIV-1 in infected individuals. In this study the COBAS® TaqMan® HIV-1 test was evaluated for use with the High Pure System (HPS), for quantifying HIV-1 RNA in infected cells and lymphoid tissue specimens. Serial dilutions of 8E5–LAV1 infected T-cells into SUP-T1 cells and 44 tonsil specimens were examined. Some modifications of the test were required, such as the removal of residual DNA and the HIV-1 RNA output copies were adjusted to the sample input and expressed as HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan HIV-1® (HPS) test proved to be a robust, sensitive, specific and reproducible method for quantifying HIV-1 RNA in infected cells and lymphoid tissue. Linearity and reproducibility were observed in serial dilutions of 8E5–LAV1 infected T-cells (R2>0.86). High reproducibility was found in clinical tonsil specimens (Wilcoxon test p>0.05). rDNase I treatment was essential to avoid false positives caused by residual HIV-1 DNA, mainly in tonsil specimens obtained from infected patients receiving effective antiretroviral treatment. Probit analysis determined the limit of detection as 22HIV-1 RNA copies/μg of total RNA. The Roche COBAS® TaqMan® HIV-1 (HPS) test thereby proved to be a helpful tool for measuring the HIV-1 viral load in infected cells and lymphoid tissue reservoirs.</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>21458494</pmid><doi>10.1016/j.jviromet.2011.03.026</doi><tpages>8</tpages></addata></record> |
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| subjects | antiretroviral agents Biological and medical sciences Cells, Cultured COBAS TaqMan HIV-1 detection limit DNA Fundamental and applied biological sciences. Psychology HIV infections HIV-1 - genetics HIV-1 - isolation & purification HIV-1 RNA quantitation Human immunodeficiency virus 1 Humans Lymphoid tissue Microbiology Nucleic Acid Amplification Techniques - methods Palatine Tonsil - virology patients probit analysis Reagent Kits, Diagnostic Reproducibility of Results RNA RNA, Viral - genetics RNA, Viral - isolation & purification Sensitivity and Specificity T-lymphocytes T-Lymphocytes - virology Techniques used in virology tonsils viral load Viral Load - methods Virology |
| title | Evaluation of the Roche COBAS® TaqMan® HIV-1 test for quantifying HIV-1 RNA in infected cells and lymphoid tissue |
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