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.
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.
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Miriam Y H
authors.initials: MY
authors.lastname: Ueda
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Paulo G
authors.initials: PG
authors.lastname: Alvarenga
authors.affiliation: Centro de Oncologia, Instituto de Ensino e Pesquisa, Hospital Sírio Libanês, São Paulo, SP, Brasil.
authors.forename: Juliana M
authors.initials: JM
authors.lastname: Real
authors.affiliation: Dendrix Research, São Paulo, SP, Brasil.
authors.forename: Eloisa de Sá
authors.initials: Ede S
authors.lastname: Moreira
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Aripuanã
authors.initials: A
authors.lastname: Watanabe
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Passos-Castilho
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Matheus
authors.initials: M
authors.lastname: Vescovi
authors.affiliation: Centro de Oncologia, Instituto de Ensino e Pesquisa, Hospital Sírio Libanês, São Paulo, SP, Brasil.
authors.forename: Yana
authors.initials: Y
authors.lastname: Novis
authors.affiliation: Centro de Oncologia, Instituto de Ensino e Pesquisa, Hospital Sírio Libanês, São Paulo, SP, Brasil.
authors.forename: Vanderson
authors.initials: V
authors.lastname: Rocha
authors.affiliation: Instituto de Oncologia Pediátrica, São Paulo, SP, Brasil.
authors.forename: Adriana
authors.initials: A
authors.lastname: Seber
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Jose S R
authors.initials: JS
authors.lastname: Oliveira
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Celso A
authors.initials: CA
authors.lastname: Rodrigues
authors.affiliation: Universidade Federal de São Paulo, São Paulo, SP, Brasil.
authors.forename: Celso F H
authors.initials: CF
authors.lastname: Granato
mesh_terms: D004279:DNA, Viral / Q000032:analysis*; D018380:Hematopoietic Stem Cell Transplantation; D015654:Herpesvirus 6, Human / Q000235:genetics*; D006801:Humans; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D019349:Roseolovirus Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D014184:Transplantation, Homologous; D019562:Viral Load
chemical_list: D004279:DNA, Viral