Molecular Quantification of Respiratory Syncytial Virus in Respiratory Samples: Reliable Detection during the Initial Phase of Infection
Quantitative real-time PCR for the detection of respiratory syncytial virus (RSV) RNA is increasingly used to study the causal role of RSV in lower airway disease. The objective of our study was to evaluate variations in RSV RNA loads at different steps in the RNA quantification process: (i) variati...
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| Veröffentlicht in: | Journal of Clinical Microbiology 2010-10, Vol.48 (10), p.3569-3574 |
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| creator | van de Pol, Alma C Wolfs, Tom F.W van Loon, Anton M Tacke, Carline E.A Viveen, Marco C Jansen, Nicolaas J.G Kimpen, Jan L.L Rossen, John W.A Coenjaerts, Frank E.J |
| description | Quantitative real-time PCR for the detection of respiratory syncytial virus (RSV) RNA is increasingly used to study the causal role of RSV in lower airway disease. The objective of our study was to evaluate variations in RSV RNA loads at different steps in the RNA quantification process: (i) variation in RSV RNA load within one sample (step 1), (ii) variation in the load in samples from patients who were sampled twice on the same day (step 2), and (iii) variation in the load between simultaneously taken nasopharyngeal aspirate (NPA) samples and tracheal aspirate (TA) samples (step 3). Thirty-two infants with RSV infection at the pediatric intensive care unit (PICU) were included. NPA and TA samples were taken three times a week during ventilation and were not diluted. Intrasample variation (step 1) was shown to be minimal ( |
| doi_str_mv | 10.1128/JCM.00097-10 |
| format | Article |
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The objective of our study was to evaluate variations in RSV RNA loads at different steps in the RNA quantification process: (i) variation in RSV RNA load within one sample (step 1), (ii) variation in the load in samples from patients who were sampled twice on the same day (step 2), and (iii) variation in the load between simultaneously taken nasopharyngeal aspirate (NPA) samples and tracheal aspirate (TA) samples (step 3). Thirty-two infants with RSV infection at the pediatric intensive care unit (PICU) were included. NPA and TA samples were taken three times a week during ventilation and were not diluted. Intrasample variation (step 1) was shown to be minimal (<0.5 log₁₀ particles/ml). Intraday variation (step 2) was the lowest for samples with high viral loads (95% limits of agreement, -1.3 to +0.9 log₁₀), whereas it increased for samples with relatively lower viral loads (viral load, <6.0 log₁₀ particles/ml; n = 138 sample pairs from 20 patients). RSV loads in NPA and TA samples (step 3) were found to be the most comparable during the early phase of infection (95% limits of agreement, -1.5 to +1.4 log₁₀). The variation increased during the late phase of infection (i.e., in follow-up samples), with the loads in NPA samples remaining significantly higher than the loads in TA samples (n = 138 sample pairs from 31 patients). In conclusion, quantitative detection of RSV RNA in undiluted mucus is a reliable method to quantify viral loads. Nasopharyngeal aspirate samples collected in the initial phase of infection can be used to predict RSV RNA loads in the lower airways.</description><identifier>ISSN: 0095-1137</identifier><identifier>EISSN: 1098-660X</identifier><identifier>DOI: 10.1128/JCM.00097-10</identifier><identifier>PMID: 20660210</identifier><identifier>CODEN: JCMIDW</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Biological and medical sciences ; Fundamental and applied biological sciences. Psychology ; Humans ; Infant ; Microbiology ; Mucus - virology ; Nasopharynx - virology ; Respiratory syncytial virus ; Respiratory Syncytial Virus Infections - virology ; Respiratory Syncytial Virus, Human - isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction - methods ; RNA, Viral - genetics ; Trachea - virology ; Viral Load ; Virology ; Virology - methods</subject><ispartof>Journal of Clinical Microbiology, 2010-10, Vol.48 (10), p.3569-3574</ispartof><rights>2015 INIST-CNRS</rights><rights>Copyright © 2010, American Society for Microbiology 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c528t-1e6a9c3d5e4854e47d3440f1de9442209eac3717b78e0f0a68af10bcf3a8d6823</citedby><cites>FETCH-LOGICAL-c528t-1e6a9c3d5e4854e47d3440f1de9442209eac3717b78e0f0a68af10bcf3a8d6823</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2953131/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2953131/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,3175,3176,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23304193$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20660210$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>van de Pol, Alma C</creatorcontrib><creatorcontrib>Wolfs, Tom F.W</creatorcontrib><creatorcontrib>van Loon, Anton M</creatorcontrib><creatorcontrib>Tacke, Carline E.A</creatorcontrib><creatorcontrib>Viveen, Marco C</creatorcontrib><creatorcontrib>Jansen, Nicolaas J.G</creatorcontrib><creatorcontrib>Kimpen, Jan L.L</creatorcontrib><creatorcontrib>Rossen, John W.A</creatorcontrib><creatorcontrib>Coenjaerts, Frank E.J</creatorcontrib><title>Molecular Quantification of Respiratory Syncytial Virus in Respiratory Samples: Reliable Detection during the Initial Phase of Infection</title><title>Journal of Clinical Microbiology</title><addtitle>J Clin Microbiol</addtitle><description>Quantitative real-time PCR for the detection of respiratory syncytial virus (RSV) RNA is increasingly used to study the causal role of RSV in lower airway disease. The objective of our study was to evaluate variations in RSV RNA loads at different steps in the RNA quantification process: (i) variation in RSV RNA load within one sample (step 1), (ii) variation in the load in samples from patients who were sampled twice on the same day (step 2), and (iii) variation in the load between simultaneously taken nasopharyngeal aspirate (NPA) samples and tracheal aspirate (TA) samples (step 3). Thirty-two infants with RSV infection at the pediatric intensive care unit (PICU) were included. NPA and TA samples were taken three times a week during ventilation and were not diluted. Intrasample variation (step 1) was shown to be minimal (<0.5 log₁₀ particles/ml). Intraday variation (step 2) was the lowest for samples with high viral loads (95% limits of agreement, -1.3 to +0.9 log₁₀), whereas it increased for samples with relatively lower viral loads (viral load, <6.0 log₁₀ particles/ml; n = 138 sample pairs from 20 patients). RSV loads in NPA and TA samples (step 3) were found to be the most comparable during the early phase of infection (95% limits of agreement, -1.5 to +1.4 log₁₀). The variation increased during the late phase of infection (i.e., in follow-up samples), with the loads in NPA samples remaining significantly higher than the loads in TA samples (n = 138 sample pairs from 31 patients). In conclusion, quantitative detection of RSV RNA in undiluted mucus is a reliable method to quantify viral loads. Nasopharyngeal aspirate samples collected in the initial phase of infection can be used to predict RSV RNA loads in the lower airways.</description><subject>Biological and medical sciences</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Infant</subject><subject>Microbiology</subject><subject>Mucus - virology</subject><subject>Nasopharynx - virology</subject><subject>Respiratory syncytial virus</subject><subject>Respiratory Syncytial Virus Infections - virology</subject><subject>Respiratory Syncytial Virus, Human - isolation & purification</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>RNA, Viral - genetics</subject><subject>Trachea - virology</subject><subject>Viral Load</subject><subject>Virology</subject><subject>Virology - methods</subject><issn>0095-1137</issn><issn>1098-660X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkkuLFDEUhYMoTtu6c62FIG6sMc-q1CwEaV8tM_gYR9yF26mkO0Mq1SZVSv8Df7bph6OzUrIInPtxTi45CN0n-JgQKp-9m50dY4ybuiT4BpoQ3MiyqvDXm2iSVVESwuojdCelS4wJ50LcRkcUZ4ISPEE_z3pv9OghFh9HCIOzTsPg-lD0tvhk0tpFGPq4Kc43QW8GB7744uKYCheuj6Fbe5NOsugdLLwpXprB6J1TO0YXlsWwMsU8uJ3HhxUks42YB7un7qJbFnwy9w73FF28fvV59rY8ff9mPntxWmpB5VASU0GjWSsMl4IbXreMc2xJaxrOKcWNAc1qUi9qabDFUEmwBC-0ZSDbSlI2Rc_3vutx0ZlWmzBE8GodXQdxo3pw6vokuJVa9t8VbQQj-UzRk4NB7L-NJg2qc0kb7yGYfkxKEiE4pkz-F8lE1TT_JGtR5Q_jnGXy6Z7UsU8pGnv1coLVtg8q90Ht-pCVjD_4e9sr-HcBMvD4AEDS4G2EoF36wzGGOWm2uY_23MotVz9cNApSpy51p7jcRm_3yNDDPWShV7CM2ejinGLCMJENrxllvwDGSdRp</recordid><startdate>20101001</startdate><enddate>20101001</enddate><creator>van de Pol, Alma C</creator><creator>Wolfs, Tom F.W</creator><creator>van Loon, Anton M</creator><creator>Tacke, Carline E.A</creator><creator>Viveen, Marco C</creator><creator>Jansen, Nicolaas J.G</creator><creator>Kimpen, Jan L.L</creator><creator>Rossen, John W.A</creator><creator>Coenjaerts, Frank E.J</creator><general>American Society for Microbiology</general><general>American Society for Microbiology (ASM)</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><scope>7U9</scope><scope>H94</scope><scope>5PM</scope></search><sort><creationdate>20101001</creationdate><title>Molecular Quantification of Respiratory Syncytial Virus in Respiratory Samples: Reliable Detection during the Initial Phase of Infection</title><author>van de Pol, Alma C ; Wolfs, Tom F.W ; van Loon, Anton M ; Tacke, Carline E.A ; Viveen, Marco C ; Jansen, Nicolaas J.G ; Kimpen, Jan L.L ; Rossen, John W.A ; Coenjaerts, Frank E.J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c528t-1e6a9c3d5e4854e47d3440f1de9442209eac3717b78e0f0a68af10bcf3a8d6823</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Biological and medical sciences</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Infant</topic><topic>Microbiology</topic><topic>Mucus - virology</topic><topic>Nasopharynx - virology</topic><topic>Respiratory syncytial virus</topic><topic>Respiratory Syncytial Virus Infections - virology</topic><topic>Respiratory Syncytial Virus, Human - isolation & purification</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>RNA, Viral - genetics</topic><topic>Trachea - virology</topic><topic>Viral Load</topic><topic>Virology</topic><topic>Virology - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>van de Pol, Alma C</creatorcontrib><creatorcontrib>Wolfs, Tom F.W</creatorcontrib><creatorcontrib>van Loon, Anton M</creatorcontrib><creatorcontrib>Tacke, Carline E.A</creatorcontrib><creatorcontrib>Viveen, Marco C</creatorcontrib><creatorcontrib>Jansen, Nicolaas J.G</creatorcontrib><creatorcontrib>Kimpen, Jan L.L</creatorcontrib><creatorcontrib>Rossen, John W.A</creatorcontrib><creatorcontrib>Coenjaerts, Frank E.J</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><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Clinical Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>van de Pol, Alma C</au><au>Wolfs, Tom F.W</au><au>van Loon, Anton M</au><au>Tacke, Carline E.A</au><au>Viveen, Marco C</au><au>Jansen, Nicolaas J.G</au><au>Kimpen, Jan L.L</au><au>Rossen, John W.A</au><au>Coenjaerts, Frank E.J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular Quantification of Respiratory Syncytial Virus in Respiratory Samples: Reliable Detection during the Initial Phase of Infection</atitle><jtitle>Journal of Clinical Microbiology</jtitle><addtitle>J Clin Microbiol</addtitle><date>2010-10-01</date><risdate>2010</risdate><volume>48</volume><issue>10</issue><spage>3569</spage><epage>3574</epage><pages>3569-3574</pages><issn>0095-1137</issn><eissn>1098-660X</eissn><coden>JCMIDW</coden><abstract>Quantitative real-time PCR for the detection of respiratory syncytial virus (RSV) RNA is increasingly used to study the causal role of RSV in lower airway disease. The objective of our study was to evaluate variations in RSV RNA loads at different steps in the RNA quantification process: (i) variation in RSV RNA load within one sample (step 1), (ii) variation in the load in samples from patients who were sampled twice on the same day (step 2), and (iii) variation in the load between simultaneously taken nasopharyngeal aspirate (NPA) samples and tracheal aspirate (TA) samples (step 3). Thirty-two infants with RSV infection at the pediatric intensive care unit (PICU) were included. NPA and TA samples were taken three times a week during ventilation and were not diluted. Intrasample variation (step 1) was shown to be minimal (<0.5 log₁₀ particles/ml). Intraday variation (step 2) was the lowest for samples with high viral loads (95% limits of agreement, -1.3 to +0.9 log₁₀), whereas it increased for samples with relatively lower viral loads (viral load, <6.0 log₁₀ particles/ml; n = 138 sample pairs from 20 patients). RSV loads in NPA and TA samples (step 3) were found to be the most comparable during the early phase of infection (95% limits of agreement, -1.5 to +1.4 log₁₀). The variation increased during the late phase of infection (i.e., in follow-up samples), with the loads in NPA samples remaining significantly higher than the loads in TA samples (n = 138 sample pairs from 31 patients). In conclusion, quantitative detection of RSV RNA in undiluted mucus is a reliable method to quantify viral loads. Nasopharyngeal aspirate samples collected in the initial phase of infection can be used to predict RSV RNA loads in the lower airways.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>20660210</pmid><doi>10.1128/JCM.00097-10</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Biological and medical sciences Fundamental and applied biological sciences. Psychology Humans Infant Microbiology Mucus - virology Nasopharynx - virology Respiratory syncytial virus Respiratory Syncytial Virus Infections - virology Respiratory Syncytial Virus, Human - isolation & purification Reverse Transcriptase Polymerase Chain Reaction - methods RNA, Viral - genetics Trachea - virology Viral Load Virology Virology - methods |
| title | Molecular Quantification of Respiratory Syncytial Virus in Respiratory Samples: Reliable Detection during the Initial Phase of Infection |
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