Highly sensitive and broadly reactive quantitative reverse transcription-PCR assay for high-throughput detection of Rift Valley fever virus

Brian H. Bird, Darcy A. Bawiec, Thomas Ksiazek, Trevor R. Shoemaker, Stuart T. Nichol

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Rift Valley fever (RVF) virus is a mosquito-borne virus associated with large-scale epizootics/epidemics throughout Africa and the Arabian peninsula. Virus infection can result in economically disastrous "abortion storms" and high newborn mortality in livestock. Human infections result in a flu-like illness, with 1 to 2% of patients developing severe complications, including encephalitis or hemorrhagic fever with high fatality rates. There is a critical need for a highly sensitive and specific molecular diagnostic assay capable of detecting the natural genetic spectrum of RVF viruses. We report here the establishment of a pan-RVF virus quantitative real-time reverse transcription-PCR assay with high analytical sensitivity (∼5 RNA copies of in vitro-transcribed RNA/reaction or ∼0.1 PFU of infectious virus/reaction) and efficiency (standard curve slope = -3.66). Based on the alignments of the complete genome sequences of 40 ecologically and biologically diverse virus isolates collected over 56 years (1944 to 2000), the primer and probe annealing sites targeted in this assay are known to be located in highly conserved genomic regions. The performance of this assay relative to serologic assays is illustrated by testing of known RVF case materials obtained during the Saudi Arabia outbreak in 2000. Furthermore, analysis of acute-phase blood samples collected from human patients (25 nonfatal, 8 fatal) during that outbreak revealed that patient viremia at time of presentation at hospital may be a useful prognostic tool in determining patient outcome.

Original languageEnglish (US)
Pages (from-to)3506-3513
Number of pages8
JournalJournal of Clinical Microbiology
Volume45
Issue number11
DOIs
StatePublished - Nov 2007
Externally publishedYes

Fingerprint

Rift Valley fever virus
Reverse Transcription
Polymerase Chain Reaction
Viruses
Disease Outbreaks
Rift Valley Fever
RNA
Molecular Pathology
Saudi Arabia
Viremia
Livestock
Virus Diseases
Encephalitis
Culicidae
Fever
Newborn Infant
Genome
Mortality
Infection

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Highly sensitive and broadly reactive quantitative reverse transcription-PCR assay for high-throughput detection of Rift Valley fever virus. / Bird, Brian H.; Bawiec, Darcy A.; Ksiazek, Thomas; Shoemaker, Trevor R.; Nichol, Stuart T.

In: Journal of Clinical Microbiology, Vol. 45, No. 11, 11.2007, p. 3506-3513.

Research output: Contribution to journalArticle

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