Journal
JOURNAL OF CLINICAL MICROBIOLOGY
Volume 50, Issue 12, Pages 4054-4060Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JCM.01799-12
Keywords
-
Categories
Funding
- Federal Ministry of Education and Research (BMBF) under the research program for civil security of the German Federal Government, as part of the high-tech strategy for potential release-oriented biothreat emergency diagnostics (P.R.O.B.E.)
Ask authors/readers for more resources
Reported methods for the detection of the yellow fever viral genome are beset by limitations in sensitivity, specificity, strain detection spectra, and suitability to laboratories with simple infrastructure in areas of endemicity. We describe the development of two different approaches affording sensitive and specific detection of the yellow fever genome: a real-time reverse transcription-quantitative PCR (RT-qPCR) and an isothermal protocol employing the same primer-probe set but based on helicase-dependent amplification technology (RT-tHDA). Both assays were evaluated using yellow fever cell culture supernatants as well as spiked and clinical samples. We demonstrate reliable detection by both assays of different strains of yellow fever virus with improved sensitivity and specificity. The RT-qPCR assay is a powerful tool for reference or diagnostic laboratories with real-time PCR capability, while the isothermal RT-tHDA assay represents a useful alternative to earlier amplification techniques for the molecular diagnosis of yellow fever by field or point-of-care laboratories.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available