Identification of genetic variation among St. Louis encephalitis virus isolates, using single-strand conformation polymorphism analysis.
A single-strand conformation polymorphism (SSCP) technique was developed for identification of genetic variation among 26 isolates of St. Louis encephalitis (SLE) virus. A 750-bp portion of the envelope gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the products analyzed by SSCP. SSCP reliably identified genetic variation among the isolates from the US, Central and South America. Closely related isolates from a smaller geographic area (Panama) were also distinguishable by SSCP. The sensitivity of this technique was demonstrated by sequencing each of the isolates used; SSCP was capable of discriminating between isolates that had as few as 1-6 nucleotide differences. These results indicate that SSCP has excellent potential as a tool to screen rapidly SLE virus isolates for genetic variation and could be incorporated into molecular epidemiology studies.
Chandler
L J
LJ
Department of Pathology and Center for Tropical Diseases, University of Texas Medical Branch, Galveston 77555, USA. laura.chandler@utmb.edu
Nordoff
N G
NG
D000818:Animals; D001103:Arboviruses / Q000235:genetics; D002519:Ceratopogonidae / Q000821:virology; D009033:Culicidae / Q000821:virology; D004665:Encephalitis Virus, St. Louis / Q000235:genetics* / Q000302:isolation & purification*; D005416:Flavivirus / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics*; D006801:Humans; D018807:Polymorphism, Single-Stranded Conformational; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity