Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2006 Aug;135(2):235-9.
doi: 10.1016/j.jviromet.2006.03.009. Epub 2006 May 2.

A single tube RT-PCR assay for the detection of mosquito-borne flaviviruses

M Ayers  1 , D Adachi, G Johnson, M Andonova, M Drebot, R Tellier
Affiliations

A single tube RT-PCR assay for the detection of mosquito-borne flaviviruses

M Ayers et al. J Virol Methods. 2006 Aug.

Abstract

Mosquito-borne flaviviruses include several important agents of human disease and have provided striking examples of emerging infections. In this study we present the design and validation of a single tube RT-PCR assay using a pair of consensus primers for the detection of mosquito-borne flaviviruses. Sequencing of the amplicons permits the species identification. The assay was validated using RNA from the yellow fever virus vaccine strain and from representative strains of dengue viruses 1, 2, 3 and 4, West Nile virus, Kunjin virus (a clade of West Nile virus), and St. Louis encephalitis virus.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Alignments calculated from the nucleotide sequences of several flavivirus species with the primer FLAVI-1 (A) or with the reverse-complemented sequence of primer FLAVI-2 (B). Dots indicate identity with the consensus sequence on top of the alignment. West Nile virus strain NY2000 (wnny): GenBank accession #AF404756; West Nile virus strain MD2000 (wnmd): #AF404753; West Nile virus strain Eg 101 (wneg 101): #AF260968; Kunjin virus: #D00246; St. Louis encephalitis virus (sle): AF013416; Murray Valley encephalitis (mve): NC_000943; Japanese encephalitis virus (jev): AF254452; Usutu virus: #AF013412; dengue virus 1 (den1): AB074766; dengue virus type 2 (den2): NC_001474; dengue virus type 3 (den3): NC_001475; dengue virus type 4 (den4): NC_002640; yellow fever virus (yfv): #YFV54798.
Fig. 2
Fig. 2
Phylogenetic tree calculated from the predicted sequences of amplicons generated by the RT-PCR assay. Bootstrap values are indicated (as percentage) above the nodes. Scale: substitution/site. WNV(NY): West Nile virus strain NY2000; WNV(Md): West Nile virus strain Md2000; WNV(Eg 101): West Nile virus strain Eg 101; JEV: Japanese encephalitis virus; MVE: Murray Valley encephalitis; SLE: St. Louis encephalitis virus; YFV: yellow fever virus.
See this image and copyright information in PMC

References

    1. Burke D.S., Monath T.P. Flaviviruses. In: Knipe D.M., Howley P.M., Griffin D.E., Lamb R.A., Martin M.A., Roizman B., editors. 4th ed. vol. 1. Lippincott Williams 7 Wilkins; Philadelphia: 2001. pp. 1043–1126. (Fields’Virology).
    1. Busch M.P., Tobler L.H., Saldanha J., Caglioti S., Shyamala V., Linnen J.M., Gallarda J., Phelps B., Smith R.I., Drebot M., Kleinman S.H. Analytical and clinical sensitivity of West Nile virus RNA screening and supplemental assays available in 2003. Transfusion. 2005;45:492–499. - PubMed
    1. CDC, 2005. West Nile virus.
    1. Davis, C.T., Ebel, G.D., Lanciotti, R.S., Brault, A.C., Guzman, H., Siirin, M., Lambert, A., Parsons, R.E., Beasley, D.W., Novak, R.J., Elizondo-Quiroga, D., Green, E.N., Young, D.S., Stark, L.M., Drebot, M.A., Artsob, H., Tesh, R.B., Kramer, L.D., Barrett, A.D., 2005. Phylogenetic analysis of North American West Nile virus isolates, 2001–2004: evidence for the emergence of a dominant genotype. Virology. - PubMed
    1. Drebot M.A., Artsob H. West Nile virus. Update for family physicians. Can. Fam. Physician. 2005;51:1094–1099. - PMC - PubMed

Publication types

MeSH terms

LinkOut - more resources