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Puumala hantavirus viremia diagnosed by real-time reverse transcriptase PCR using samples from patients with hemorrhagic fever and renal syndrome.

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  • Additional Information
    • Publisher Information:
      Umeå universitet, Virologi Umeå universitet, Virologi Umeå universitet, Virologi Umeå universitet, Virologi Umeå universitet, Infektionssjukdomar Umeå universitet, Virologi Umeå universitet, Institutionen för molekylärbiologi (Teknisk-naturvetenskaplig fakultet) Umeå universitet, Virologi 2007
    • Abstract:
      Puumala virus (PUUV) is the endemic hantavirus in northern Sweden and causes nephropathia epidemica (NE), a milder form of hemorrhagic fever with renal syndrome. There is a need for fast and reliable diagnostics to differentiate the disease from other infections. By aligning virus RNA sequences isolated from 11 different bank voles and one human patient, we designed a real-time reverse transcriptase (RT) PCR method for detection of PUUV RNA. The real-time RT-PCR assay showed linearity from 20 to 2 x 10(6) virus copies with a correlation coefficient above 0.98 to 0.99 for all experiments. The detection threshold for PUUV cDNA was two copies per reaction. A two-step qualitative RT-PCR to detect PUUV RNA showed 100% concordance with the real-time RT-PCR assay. PUUV RNA viremia was detected in 33 of 34 PUUV immunoglobulin M (IgM)-positive patients with typical clinical NE disease from the region of endemicity. One PUUV IgM-negative sample had PUUV RNA, and 4 days later, the patient was IgM positive. Of samples with indeterminate IgM, 43% were PUUV RNA positive. The kinetics of antibody titers and PUUV viremia were studied, and five of six NE patients displayed a decrease in PUUV viremia a few days after disease outbreak coupled with an increase in PUUV IgM and IgG. In one patient with continuously high PUUV RNA levels but low IgM and no IgG response, the infection was lethal. These findings demonstrated that real-time RT-PCR is a useful method for diagnosis of PUUV viremia and for detecting PUUV RNA at early time points, before the appearance of IgM antibodies.
    • Subject Terms:
      Article in journal; info:eu-repo/semantics/article; text
    • Accession Number:
      10.1128.JCM.01902-06
    • Availability:
      Open access content. Open access content
      info:eu-repo/semantics/restrictedAccess
    • Note:
      English
    • Other Numbers:
      UPE oai:DiVA.org:umu-20652
      doi:10.1128/JCM.01902-06
      PMID 17537944
      1234313457
    • Contributing Source:
      UPPSALA UNIV LIBR
      From OAIster®, provided by the OCLC Cooperative.
    • Accession Number:
      edsoai.on1234313457
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