Rapid identification of dengue virus by reverse transcription-polymerase chain reaction using field-deployable instrumentation.
Dengue virus universal and dengue serotype 1 to 4, fluorogenic probe hydrolysis (TaqMan), reverse transcription-polymerase chain reaction assays were developed for screening and serotype identification of infected mosquito vectors and human sera using a field-deployable, fluorometric thermocycler. Dengue universal and dengue 1 to 4 serotype assay in vitro sensitivity and specificity results were 100% concordant when tested with total nucleic acid extracts of multiple strains of dengue serotype 1 to 4, yellow fever, Japanese encephalitis, West Nile, and St. Louis encephalitis viruses. The in vitro sensitivity and specificity results for all five assays were concordant when tested with a blind panel of 27 dengue virus-infected mosquitoes, 21 non-dengue (yellow fever, West Nile, or St. Louis encephalitis) flavivirus-infected mosquitoes, and 11 uninfected mosquitoes and with clinical specimens consisting of a human serum panel of eight dengue viremic and 31 non-dengue-infected febrile patient serum samples. No cross-reaction occurred with vector species or human genomic DNA. Sample processing and polymerase chain reaction required <2 hours.
Epidemiological Surveillance Division, U.S. Air Force Institute for Environment, Safety, and Occupational Health Analysis, Brooks Air Force Base (now designated Air Force Institute for Operational Health, Brooks City-Base), San Antonio, TX 78235-5237, USA.
James C
JC
McAvin
Elizabeth M
EM
Escamilla
Jamie A
JA
Blow
Michael J
MJ
Turell
Miguel
M
Quintana
David E
DE
Bowles
James A
JA
Swaby
William J
WJ
Barnes
William B
WB
Huff
Kenton L
KL
Lohman
Daniel H
DH
Atchley
John R
JR
Hickman
Debra M
DM
Niemeyer
D000818:Animals; D009033:Culicidae / Q000235:genetics* / Q000821:virology; D003715:Dengue / Q000097:blood / Q000821:virology*; D003716:Dengue Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007303:Insect Vectors / Q000821:virology*; D008887:Military Medicine / Q000295:instrumentation*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation*