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

Memórias do Instituto Oswaldo Cruz

Miriam Yh Ueda¹  Paulo G Alvarenga¹  Juliana M Real¹  Eloisa De Sá Moreira¹  Aripuanã Watanabe¹  Ana Maria Passos-castilho¹  Matheus Vescovi¹  Yana Novis¹  Vanderson Rocha¹  Adriana Seber¹  Jose Sr Oliveira¹  Celso A Rodrigues¹  Celso Fh Granato¹ 

关键词: human herpesvirus 6;    real-time PCR;    viral load;   

DOI  :  10.1590/0074-02760150004

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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