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
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 incide...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2015-06-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461&lng=en&tlng=en |
| _version_ | 1797757585293574144 |
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| author | Miriam YH Ueda Paulo G Alvarenga Juliana M Real Eloisa de Sá Moreira Aripuanã Watanabe Ana Maria Passos-Castilho Matheus Vescovi Yana Novis Vanderson Rocha Adriana Seber Jose SR Oliveira Celso A Rodrigues Celso FH Granato |
| author_facet | Miriam YH Ueda Paulo G Alvarenga Juliana M Real Eloisa de Sá Moreira Aripuanã Watanabe Ana Maria Passos-Castilho Matheus Vescovi Yana Novis Vanderson Rocha Adriana Seber Jose SR Oliveira Celso A Rodrigues Celso FH Granato |
| author_sort | Miriam YH Ueda |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-12T18:17:57Z |
| format | Article |
| id | doaj.art-2063b76a0ac3481698512aa9307424e2 |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T18:17:57Z |
| publishDate | 2015-06-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-2063b76a0ac3481698512aa9307424e22023-08-02T09:00:11ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-80602015-06-01110446146710.1590/0074-02760150004S0074-02762015000400461Optimisation 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 transplantationMiriam YH UedaPaulo G AlvarengaJuliana M RealEloisa de Sá MoreiraAripuanã WatanabeAna Maria Passos-CastilhoMatheus VescoviYana NovisVanderson RochaAdriana SeberJose SR OliveiraCelso A RodriguesCelso FH GranatoHuman 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461&lng=en&tlng=enhuman herpesvirus 6real-time PCRviral load |
| spellingShingle | Miriam YH Ueda Paulo G Alvarenga Juliana M Real Eloisa de Sá Moreira Aripuanã Watanabe Ana Maria Passos-Castilho Matheus Vescovi Yana Novis Vanderson Rocha Adriana Seber Jose SR Oliveira Celso A Rodrigues Celso FH Granato 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 Memorias do Instituto Oswaldo Cruz human herpesvirus 6 real-time PCR viral load |
| 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 |
| 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_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_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 |
| topic | human herpesvirus 6 real-time PCR viral load |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400461&lng=en&tlng=en |
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