Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus.
Transmission experiments are a critical component of vector competence studies. In this study, a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to enumerate the amount of West Nile virus (WNV) secreted in mosquito saliva following oral infection. Culex pipiens quinquefasciatus were allowed to feed on WNV-infected blood, and saliva was collected on days 14 and 21 post-infection (pi). The amount of virus at these two time points varied significantly, with mean equivalent plaque-forming units (pfu) of approximately 30,500 on day 14 pi and 5,800 on day 21 pi. Individual mosquitoes secreted up to 2 x 10(5) pfu of virus. Titer of whole mosquitoes and immunofluorescence assay of salivary glands from mosquitoes collected at these two time points were also used for supplemental comparison. This report describes the first use of a real-time RT-PCR to quantify the amount of WNV in mosquito saliva.
Vanlandingham
Dana L
DL
Department of Pathology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA.
Schneider
Bradley S
BS
Klingler
Kimberly
K
Fair
Joseph
J
Beasley
David
D
Huang
Jing
J
Hamilton
Patricia
P
Higgs
Stephen
S
D000818:Animals; D000956:Antigens, Viral; D003465:Culex / Q000821:virology*; D007303:Insect Vectors / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014901:West Nile Fever / Q000453:epidemiology / Q000635:transmission* / Q000821:virology; D014902:West Nile virus / Q000235:genetics / Q000502:physiology*
D000956:Antigens, Viral