Reliable detection of St. Louis encephalitis virus by RT-nested PCR

RÉ, VIVIANA ELIZABETH; SPINSANTI, LORENA IVANA; FARÍAS, ADRIÁN ALEJANDRO; DIAZ, LUIS ADRIÁN; VÁZQUEZ, ANA; AGUILAR, JAVIER JOSÉ; TENORIO, ANTONIO; CONTIGIANI, MARTA SILVIA

ENFERMEDADES INFECCIOSAS Y MICROBIOLOGIA CLINICA.

Ediciones Doyma, SL - Elsevier

Lugar: Madrid, España; Año: 2008 vol. 26 p. 10 - 15

0213-005X

We report the development of a species-specific RT-nested PCR to detect SLEV. After selecting the SLEV genomic region providing the greatest information on the natural genetic variability of this virus, degenerated oligonucleotide primers were designed to amplify a 234-bp fragment of the envelope gene from nine SLEV strains (Parton, BeH356964, SPAN11916, AN9275, AN9124, 78V6507 and 3 SLEV strains obtained from naturally infected mosquito pools). The method was able to identify the genome of all the SLEV strains tested and did not amplify unrelated RNA viruses, such as yellow fever virus, Ilheus virus, dengue-2 virus, Bussuquara virus, West Nile virus, Japanese encephalitis virus and Murray Valley encephalitis virus. The method was specific and sensitive, with a lower detection limit of < 10 plaque-forming units. This molecular assay is a reliable procedure with a wide spectrum for detecting the natural diversity of SLEV and may be useful for ecological studies, clinical and laboratory settings and virological surveillance.