Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2015 |
| Outros Autores: | , , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Memórias do Instituto Oswaldo Cruz |
| Texto Completo: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461 |
Resumo: | Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance. |
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Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantationhuman herpesvirus 6real-time PCRviral loadHuman herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.Instituto Oswaldo Cruz, Ministério da Saúde2015-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461Memórias do Instituto Oswaldo Cruz v.110 n.4 2015reponame:Memórias do Instituto Oswaldo Cruzinstname:Fundação Oswaldo Cruzinstacron:FIOCRUZ10.1590/0074-02760150004info:eu-repo/semantics/openAccessUeda,Miriam YHAlvarenga,Paulo GReal,Juliana MMoreira,Eloisa de SáWatanabe,AripuanãPassos-Castilho,Ana MariaVescovi,MatheusNovis,YanaRocha,VandersonSeber,AdrianaOliveira,Jose SRRodrigues,Celso AGranato,Celso FHeng2020-04-25T17:52:14Zhttp://www.scielo.br/oai/scielo-oai.php0074-02761678-8060opendoar:null2020-04-26 02:20:47.25Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruztrue |
| dc.title.none.fl_str_mv |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| title |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| spellingShingle |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation Ueda,Miriam YH human herpesvirus 6 real-time PCR viral load |
| title_short |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| title_full |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| title_fullStr |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| title_full_unstemmed |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| title_sort |
Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation |
| author |
Ueda,Miriam YH |
| author_facet |
Ueda,Miriam YH Alvarenga,Paulo G Real,Juliana M Moreira,Eloisa de Sá Watanabe,Aripuanã Passos-Castilho,Ana Maria Vescovi,Matheus Novis,Yana Rocha,Vanderson Seber,Adriana Oliveira,Jose SR Rodrigues,Celso A Granato,Celso FH |
| author_role |
author |
| author2 |
Alvarenga,Paulo G Real,Juliana M Moreira,Eloisa de Sá Watanabe,Aripuanã Passos-Castilho,Ana Maria Vescovi,Matheus Novis,Yana Rocha,Vanderson Seber,Adriana Oliveira,Jose SR Rodrigues,Celso A Granato,Celso FH |
| author2_role |
author author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Ueda,Miriam YH Alvarenga,Paulo G Real,Juliana M Moreira,Eloisa de Sá Watanabe,Aripuanã Passos-Castilho,Ana Maria Vescovi,Matheus Novis,Yana Rocha,Vanderson Seber,Adriana Oliveira,Jose SR Rodrigues,Celso A Granato,Celso FH |
| dc.subject.por.fl_str_mv |
human herpesvirus 6 real-time PCR viral load |
| topic |
human herpesvirus 6 real-time PCR viral load |
| dc.description.none.fl_txt_mv |
Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance. |
| description |
Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance. |
| publishDate |
2015 |
| dc.date.none.fl_str_mv |
2015-06-01 |
| dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461 |
| url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
10.1590/0074-02760150004 |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
text/html |
| dc.publisher.none.fl_str_mv |
Instituto Oswaldo Cruz, Ministério da Saúde |
| publisher.none.fl_str_mv |
Instituto Oswaldo Cruz, Ministério da Saúde |
| dc.source.none.fl_str_mv |
Memórias do Instituto Oswaldo Cruz v.110 n.4 2015 reponame:Memórias do Instituto Oswaldo Cruz instname:Fundação Oswaldo Cruz instacron:FIOCRUZ |
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Fundação Oswaldo Cruz |
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FIOCRUZ |
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FIOCRUZ |
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Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruz |
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