Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus

Dana L. Vanlandingham, Bradley S. Schneider, Kimberly Klingler, Joseph Fair, David Beasley, Jing Huang, Patricia Hamilton, Stephen Higgs

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

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 × 105 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.

Original languageEnglish (US)
Pages (from-to)120-123
Number of pages4
JournalAmerican Journal of Tropical Medicine and Hygiene
Volume71
Issue number1
StatePublished - Jul 2004

Fingerprint

West Nile virus
Culex
Culicidae
Reverse Transcriptase Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Saliva
Infection
Viruses
Salivary Glands
Mental Competency
Fluorescent Antibody Technique

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus. / Vanlandingham, Dana L.; Schneider, Bradley S.; Klingler, Kimberly; Fair, Joseph; Beasley, David; Huang, Jing; Hamilton, Patricia; Higgs, Stephen.

In: American Journal of Tropical Medicine and Hygiene, Vol. 71, No. 1, 07.2004, p. 120-123.

Research output: Contribution to journalArticle

Vanlandingham, Dana L. ; Schneider, Bradley S. ; Klingler, Kimberly ; Fair, Joseph ; Beasley, David ; Huang, Jing ; Hamilton, Patricia ; Higgs, Stephen. / Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus. In: American Journal of Tropical Medicine and Hygiene. 2004 ; Vol. 71, No. 1. pp. 120-123.
@article{89558e8124df4672be35cb5bf3ef7034,
title = "Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus",
abstract = "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 × 105 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.",
author = "Vanlandingham, {Dana L.} and Schneider, {Bradley S.} and Kimberly Klingler and Joseph Fair and David Beasley and Jing Huang and Patricia Hamilton and Stephen Higgs",
year = "2004",
month = "7",
language = "English (US)",
volume = "71",
pages = "120--123",
journal = "American Journal of Tropical Medicine and Hygiene",
issn = "0002-9637",
publisher = "American Society of Tropical Medicine and Hygiene",
number = "1",

}

TY - JOUR

T1 - Real-time reverse transcriptase-polymerase chain reaction quantification of West Nile virus transmitted by Culex pipiens quinquefasciatus

AU - Vanlandingham, Dana L.

AU - Schneider, Bradley S.

AU - Klingler, Kimberly

AU - Fair, Joseph

AU - Beasley, David

AU - Huang, Jing

AU - Hamilton, Patricia

AU - Higgs, Stephen

PY - 2004/7

Y1 - 2004/7

N2 - 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 × 105 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.

AB - 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 × 105 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.

UR - http://www.scopus.com/inward/record.url?scp=3142615274&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3142615274&partnerID=8YFLogxK

M3 - Article

C2 - 15238700

AN - SCOPUS:3142615274

VL - 71

SP - 120

EP - 123

JO - American Journal of Tropical Medicine and Hygiene

JF - American Journal of Tropical Medicine and Hygiene

SN - 0002-9637

IS - 1

ER -