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 |
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Authors: | Miriam YH Ueda Paulo G Alvarenga Juliana M Real Eloisa de Sá Moreira Aripuan? Watanabe Ana Maria Passos-Castilho Matheus Vescovi Yana Novis Vanderson Rocha Adriana Seber Jose SR Oliveira Celso A Rodrigues Celso FH Granato |
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Institution: | 1.Disciplina de Doenças Infecciosas e Parasitárias;2.Disciplina de Hematologia e Hemoterapia, Universidade Federal de São Paulo, São Paulo, SP, Brasil;3.Centro de Oncologia, Instituto de Ensino e Pesquisa, Hospital Sírio Libanês, São Paulo, SP, Brasil;4.Dendrix Research, São Paulo, SP, Brasil;5.Instituto de Oncologia Pediátrica, São Paulo, SP, Brasil |
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Abstract: | 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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Keywords: | human herpesvirus 6 real-time PCR viral load |
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