Quantification of cytomegalovirus DNA by a fully automated real-time PCR for early diagnosis and monitoring of active viral infection in solid organ transplant recipients
Abstract Background Quantification of cytomegalovirus (CMV) DNA by real-time PCR is currently considered an alternative diagnostic approach for the evaluation of active infection in transplant patients. The pp65 antigenemia assay has been used as reference test for monitoring active CMV infection an...
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| Veröffentlicht in: | Journal of clinical virology 2013-02, Vol.56 (2), p.124-128 |
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| description | Abstract Background Quantification of cytomegalovirus (CMV) DNA by real-time PCR is currently considered an alternative diagnostic approach for the evaluation of active infection in transplant patients. The pp65 antigenemia assay has been used as reference test for monitoring active CMV infection and guiding preemptive therapy in transplant recipients. However, this assay suffers from some limitations: need for immediate processing of the samples, labour-intensive process, lack of standardization and subjective result interpretation. Objectives The aim of this study was to evaluate the performance of a new commercially available real-time PCR assay coupled with a fully automated DNA extraction system (COBAS Ampliprep/COBAS Taqman CMV Test, Roche Diagnostics) for the detection of CMV-DNA in plasma comparing it with pp65 antigenemia assay for monitoring active CMV infection in solid organ transplant recipients (SOTRs). Study design A total of 266 consecutive samples from 45 SOTRs were monitored with pp65 antigenemia and in parallel with CMV-DNA quantitation by real-time PCR assay. Results Fifty-eight samples resulted PCR-positive, 163 negative and for 45 samples the CMV-DNA values obtained were below the lower limit of quantification ( |
| doi_str_mv | 10.1016/j.jcv.2012.10.015 |
| format | Article |
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The pp65 antigenemia assay has been used as reference test for monitoring active CMV infection and guiding preemptive therapy in transplant recipients. However, this assay suffers from some limitations: need for immediate processing of the samples, labour-intensive process, lack of standardization and subjective result interpretation. Objectives The aim of this study was to evaluate the performance of a new commercially available real-time PCR assay coupled with a fully automated DNA extraction system (COBAS Ampliprep/COBAS Taqman CMV Test, Roche Diagnostics) for the detection of CMV-DNA in plasma comparing it with pp65 antigenemia assay for monitoring active CMV infection in solid organ transplant recipients (SOTRs). Study design A total of 266 consecutive samples from 45 SOTRs were monitored with pp65 antigenemia and in parallel with CMV-DNA quantitation by real-time PCR assay. Results Fifty-eight samples resulted PCR-positive, 163 negative and for 45 samples the CMV-DNA values obtained were below the lower limit of quantification (<150 copies/ml); pp65 antigen was detected in 47 samples and resulted negative in 219 specimens. Concordance between the two evaluations was 76.7%; also a good correlation was observed ( r = 0.718). Considering the existing treatment criteria based on pp65 antigenemia evaluation corresponding to pp65 levels ≥ 20 positive cells/200,000, preemptive therapy was administered to four asymptomatically infected patients. The corresponding cut-off value of CMV-DNA load calculated for discrimination between self-clearing infections and those requiring therapy was 2500 copies/ml (or 2275 IU/ml). Conclusion The fully automated real-time PCR from Roche provided specific and sensitive results and represented a rapid and simple assay for the evaluation and monitoring of CMV infection in SOTRs. Further studies are required to validate the threshold level for the initiation of preemptive therapy.</description><identifier>ISSN: 1386-6532</identifier><identifier>EISSN: 1873-5967</identifier><identifier>DOI: 10.1016/j.jcv.2012.10.015</identifier><identifier>PMID: 23182772</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Adult ; Aged ; Allergy and Immunology ; Antigenemia ; Automation - methods ; Biological and medical sciences ; CMV-DNA ; Cytomegalovirus ; Cytomegalovirus Infections - diagnosis ; Cytomegalovirus Infections - virology ; DNA, Viral - isolation & purification ; Early Diagnosis ; Female ; Fundamental and applied biological sciences. Psychology ; Human viral diseases ; Humans ; Infection ; Infectious Disease ; Infectious diseases ; Male ; Medical sciences ; Microbiology ; Middle Aged ; Miscellaneous ; Polymerase chain reaction ; pp65 protein ; Quantitation ; Real-time PCR ; Real-Time Polymerase Chain Reaction - methods ; Sensitivity and Specificity ; Specimen Handling - methods ; Standardization ; Transplantation ; Transplants - adverse effects ; Viral diseases ; Viral Load - methods ; Virology ; Young Adult</subject><ispartof>Journal of clinical virology, 2013-02, Vol.56 (2), p.124-128</ispartof><rights>Elsevier B.V.</rights><rights>2012 Elsevier B.V.</rights><rights>2014 INIST-CNRS</rights><rights>Copyright © 2012 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c471t-5b55e6a5cc0fddbf4c8db4ed312e8d2700be309e9ed17d21f720c4917bd279c3</citedby><cites>FETCH-LOGICAL-c471t-5b55e6a5cc0fddbf4c8db4ed312e8d2700be309e9ed17d21f720c4917bd279c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1386653212003964$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26830869$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23182772$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Boaretti, M</creatorcontrib><creatorcontrib>Sorrentino, A</creatorcontrib><creatorcontrib>Zantedeschi, C</creatorcontrib><creatorcontrib>Forni, A</creatorcontrib><creatorcontrib>Boschiero, L</creatorcontrib><creatorcontrib>Fontana, R</creatorcontrib><title>Quantification of cytomegalovirus DNA by a fully automated real-time PCR for early diagnosis and monitoring of active viral infection in solid organ transplant recipients</title><title>Journal of clinical virology</title><addtitle>J Clin Virol</addtitle><description>Abstract Background Quantification of cytomegalovirus (CMV) DNA by real-time PCR is currently considered an alternative diagnostic approach for the evaluation of active infection in transplant patients. The pp65 antigenemia assay has been used as reference test for monitoring active CMV infection and guiding preemptive therapy in transplant recipients. However, this assay suffers from some limitations: need for immediate processing of the samples, labour-intensive process, lack of standardization and subjective result interpretation. Objectives The aim of this study was to evaluate the performance of a new commercially available real-time PCR assay coupled with a fully automated DNA extraction system (COBAS Ampliprep/COBAS Taqman CMV Test, Roche Diagnostics) for the detection of CMV-DNA in plasma comparing it with pp65 antigenemia assay for monitoring active CMV infection in solid organ transplant recipients (SOTRs). Study design A total of 266 consecutive samples from 45 SOTRs were monitored with pp65 antigenemia and in parallel with CMV-DNA quantitation by real-time PCR assay. Results Fifty-eight samples resulted PCR-positive, 163 negative and for 45 samples the CMV-DNA values obtained were below the lower limit of quantification (<150 copies/ml); pp65 antigen was detected in 47 samples and resulted negative in 219 specimens. Concordance between the two evaluations was 76.7%; also a good correlation was observed ( r = 0.718). Considering the existing treatment criteria based on pp65 antigenemia evaluation corresponding to pp65 levels ≥ 20 positive cells/200,000, preemptive therapy was administered to four asymptomatically infected patients. The corresponding cut-off value of CMV-DNA load calculated for discrimination between self-clearing infections and those requiring therapy was 2500 copies/ml (or 2275 IU/ml). Conclusion The fully automated real-time PCR from Roche provided specific and sensitive results and represented a rapid and simple assay for the evaluation and monitoring of CMV infection in SOTRs. Further studies are required to validate the threshold level for the initiation of preemptive therapy.</description><subject>Adult</subject><subject>Aged</subject><subject>Allergy and Immunology</subject><subject>Antigenemia</subject><subject>Automation - methods</subject><subject>Biological and medical sciences</subject><subject>CMV-DNA</subject><subject>Cytomegalovirus</subject><subject>Cytomegalovirus Infections - diagnosis</subject><subject>Cytomegalovirus Infections - virology</subject><subject>DNA, Viral - isolation & purification</subject><subject>Early Diagnosis</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Human viral diseases</subject><subject>Humans</subject><subject>Infection</subject><subject>Infectious Disease</subject><subject>Infectious diseases</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Microbiology</subject><subject>Middle Aged</subject><subject>Miscellaneous</subject><subject>Polymerase chain reaction</subject><subject>pp65 protein</subject><subject>Quantitation</subject><subject>Real-time PCR</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Sensitivity and Specificity</subject><subject>Specimen Handling - methods</subject><subject>Standardization</subject><subject>Transplantation</subject><subject>Transplants - adverse effects</subject><subject>Viral diseases</subject><subject>Viral Load - methods</subject><subject>Virology</subject><subject>Young Adult</subject><issn>1386-6532</issn><issn>1873-5967</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNktuKFDEQhhtR3HX0AbyR3Aje9JhDHxGEZTzC4nHvQzqpHjKmk9kkPTCv5FNa7YwKXohXOdRXfxX1V1E8ZnTNKGue79Y7fVhzyji-15TVd4pL1rWirPumvYt30TVlUwt-UTxIaUeREFV7v7jggnW8bfll8f3zrHy2o9Uq2-BJGIk-5jDBVrlwsHFO5NWHKzIciSLj7ByeM4ZVBkMiKFdmOwH5tPlCxhAJqIiEsWrrQ7KJKG_IFLzNIVq_XcSVzvYABJWVI9aPoH-WtZ6k4KwhIW6VJzkqn_YOO8Mi2u4t-JweFvdG5RI8Op-r4ubN65vNu_L649v3m6vrUlcty2U91DU0qtaajsYMY6U7M1RgBOPQGd5SOoCgPfRgWGs4G1tOddWzdsBgr8WqeHaS3cdwO0PKcrJJg8NuIMxJMt6KmjZ9xf8HRbjqBEWUnVAdQ0oRRrmPdlLxKBmVi5lyJ9FMuZi5fC1WrYonZ_l5mMD8zvjlHgJPz4BKWrkRp6Zt-sM1WLlreuRenDjAsR0sRJk0jlSDsTjeLE2w_2zj5V_Z2lmPC-O-wRHSLszRox-SycQllV-XrVuWjnFKRd9U4geNb9UM</recordid><startdate>20130201</startdate><enddate>20130201</enddate><creator>Boaretti, M</creator><creator>Sorrentino, A</creator><creator>Zantedeschi, C</creator><creator>Forni, A</creator><creator>Boschiero, L</creator><creator>Fontana, R</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>7TM</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20130201</creationdate><title>Quantification of cytomegalovirus DNA by a fully automated real-time PCR for early diagnosis and monitoring of active viral infection in solid organ transplant recipients</title><author>Boaretti, M ; Sorrentino, A ; Zantedeschi, C ; Forni, A ; Boschiero, L ; Fontana, R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c471t-5b55e6a5cc0fddbf4c8db4ed312e8d2700be309e9ed17d21f720c4917bd279c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Adult</topic><topic>Aged</topic><topic>Allergy and Immunology</topic><topic>Antigenemia</topic><topic>Automation - methods</topic><topic>Biological and medical sciences</topic><topic>CMV-DNA</topic><topic>Cytomegalovirus</topic><topic>Cytomegalovirus Infections - diagnosis</topic><topic>Cytomegalovirus Infections - virology</topic><topic>DNA, Viral - isolation & purification</topic><topic>Early Diagnosis</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Human viral diseases</topic><topic>Humans</topic><topic>Infection</topic><topic>Infectious Disease</topic><topic>Infectious diseases</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Microbiology</topic><topic>Middle Aged</topic><topic>Miscellaneous</topic><topic>Polymerase chain reaction</topic><topic>pp65 protein</topic><topic>Quantitation</topic><topic>Real-time PCR</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Sensitivity and Specificity</topic><topic>Specimen Handling - methods</topic><topic>Standardization</topic><topic>Transplantation</topic><topic>Transplants - adverse effects</topic><topic>Viral diseases</topic><topic>Viral Load - methods</topic><topic>Virology</topic><topic>Young Adult</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Boaretti, M</creatorcontrib><creatorcontrib>Sorrentino, A</creatorcontrib><creatorcontrib>Zantedeschi, C</creatorcontrib><creatorcontrib>Forni, A</creatorcontrib><creatorcontrib>Boschiero, L</creatorcontrib><creatorcontrib>Fontana, R</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>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of clinical virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Boaretti, M</au><au>Sorrentino, A</au><au>Zantedeschi, C</au><au>Forni, A</au><au>Boschiero, L</au><au>Fontana, R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of cytomegalovirus DNA by a fully automated real-time PCR for early diagnosis and monitoring of active viral infection in solid organ transplant recipients</atitle><jtitle>Journal of clinical virology</jtitle><addtitle>J Clin Virol</addtitle><date>2013-02-01</date><risdate>2013</risdate><volume>56</volume><issue>2</issue><spage>124</spage><epage>128</epage><pages>124-128</pages><issn>1386-6532</issn><eissn>1873-5967</eissn><abstract>Abstract Background Quantification of cytomegalovirus (CMV) DNA by real-time PCR is currently considered an alternative diagnostic approach for the evaluation of active infection in transplant patients. The pp65 antigenemia assay has been used as reference test for monitoring active CMV infection and guiding preemptive therapy in transplant recipients. However, this assay suffers from some limitations: need for immediate processing of the samples, labour-intensive process, lack of standardization and subjective result interpretation. Objectives The aim of this study was to evaluate the performance of a new commercially available real-time PCR assay coupled with a fully automated DNA extraction system (COBAS Ampliprep/COBAS Taqman CMV Test, Roche Diagnostics) for the detection of CMV-DNA in plasma comparing it with pp65 antigenemia assay for monitoring active CMV infection in solid organ transplant recipients (SOTRs). Study design A total of 266 consecutive samples from 45 SOTRs were monitored with pp65 antigenemia and in parallel with CMV-DNA quantitation by real-time PCR assay. Results Fifty-eight samples resulted PCR-positive, 163 negative and for 45 samples the CMV-DNA values obtained were below the lower limit of quantification (<150 copies/ml); pp65 antigen was detected in 47 samples and resulted negative in 219 specimens. Concordance between the two evaluations was 76.7%; also a good correlation was observed ( r = 0.718). Considering the existing treatment criteria based on pp65 antigenemia evaluation corresponding to pp65 levels ≥ 20 positive cells/200,000, preemptive therapy was administered to four asymptomatically infected patients. The corresponding cut-off value of CMV-DNA load calculated for discrimination between self-clearing infections and those requiring therapy was 2500 copies/ml (or 2275 IU/ml). Conclusion The fully automated real-time PCR from Roche provided specific and sensitive results and represented a rapid and simple assay for the evaluation and monitoring of CMV infection in SOTRs. Further studies are required to validate the threshold level for the initiation of preemptive therapy.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>23182772</pmid><doi>10.1016/j.jcv.2012.10.015</doi><tpages>5</tpages></addata></record> |
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| subjects | Adult Aged Allergy and Immunology Antigenemia Automation - methods Biological and medical sciences CMV-DNA Cytomegalovirus Cytomegalovirus Infections - diagnosis Cytomegalovirus Infections - virology DNA, Viral - isolation & purification Early Diagnosis Female Fundamental and applied biological sciences. Psychology Human viral diseases Humans Infection Infectious Disease Infectious diseases Male Medical sciences Microbiology Middle Aged Miscellaneous Polymerase chain reaction pp65 protein Quantitation Real-time PCR Real-Time Polymerase Chain Reaction - methods Sensitivity and Specificity Specimen Handling - methods Standardization Transplantation Transplants - adverse effects Viral diseases Viral Load - methods Virology Young Adult |
| title | Quantification of cytomegalovirus DNA by a fully automated real-time PCR for early diagnosis and monitoring of active viral infection in solid organ transplant recipients |
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