Flavivirus detection and quantification assay

摘要

Fluorescent DNA probes specific and flanking primer pairs are designed based on the sequence information found in the conserved terminal 3'-noncoding region of flavivirus, e.g. nucleotides 10653-10678 of dengue virus. Fluorogenic polymerase chain reaction employing these primers and probes produce results that permit specific flavivirus identification. The assays can be both quantitative and qualitative. Optimal assay conditions with zero background are disclosed which permit the detection of low levels of flavivirus from clinical specimens. Specifically, Dengue virus isolates from different geographic regions can be universally detected and identified by the disclosed fluorogenic RT-PCR assay. The fluorogenic RT-PCR assay readily detected viremia in sera collected from individuals ill with dengue fever.