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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060 EISSN: 1678-8060
Vol. 110, No. 4, 2015, pp. 461-467
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Bioline Code: oc15058
Full paper language: English
Document type: Research Article
Document available free of charge
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Memórias do Instituto Oswaldo Cruz, Vol. 110, No. 4, 2015, pp. 461-467
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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 transplantation
Ueda, Miriam Y.H.; 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 S.R.; Rodrigues, Celso A. & Granato, Celso F.H.
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 Taq-
Man® 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.
Keywords
human herpesvirus 6; real-time PCR; viral load
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