• 1

    Nash D, Mostashari F, Fine A, Miller J, O’Leary D, Murray K, Huang A, Rosenberg A, Greenberg A, Sherman M, Wong S, Layton M, 2001. The outbreak of West Nile virus infection in the New York City area in 1999. N Engl J Med 344 :1807–1814.

    • Search Google Scholar
    • Export Citation
  • 2

    Nasci RS, Savage HM, White DJ, Miller JR, Cropp BC, Godsey MS, Kerst AJ, Bennett P, Gottfried K, Lanciotti RS, 2001. West Nile virus in overwintering Culex mosquitoes, New York City, 2000. Emerg Infect Dis 7 :742–744.

    • Search Google Scholar
    • Export Citation
  • 3

    Kilpatrick AM, Kramer LD, Campbell SR, Alleyne EO, Dobson AP, Daszak P, 2005. West Nile virus risk assessment and the bridge vector paradigm. Emerg Infect Dis 11 :425–429.

    • Search Google Scholar
    • Export Citation
  • 4

    Sundararaman S, 1949. Biometrical studies on intergradation in the genitalia of certain populations of Culex pipiens and Culex quinquefasciatus in the United States. Am J Hyg 50 :307–314.

    • Search Google Scholar
    • Export Citation
  • 5

    Cornel AJ, McAbee RD, Rasgon J, Stanich MA, Scott TW, Coetzee M, 2003. Differences in extent of genetic introgression between sympatric Culex pipiens and Culex quinquefasciatus (Diptera: Culicidae) in California and South Africa. J Med Entomol 40 :36–51.

    • Search Google Scholar
    • Export Citation
  • 6

    Tabachnick WJ, Powell JR, 1983. Genetic analysis of Culex pipiens populations in the Central Valley of California. Ann Entomol Soc Am 76 :715–720.

    • Search Google Scholar
    • Export Citation
  • 7

    Urbanelli S, Silvestrini F, Reisen WK, DeVito E, Bullini L, 1997. Californian hybrid zone between Culex pipiens pipiens and Cx. p. quinquefasciatus revisited (Diptera: Culicidae). J Med Entomol 34 :116–127.

    • Search Google Scholar
    • Export Citation
  • 8

    Crabtree MB, Savage HM, Miller BR, 1995. Development of a species-diagnostic polymerase chain reaction assay for the identification of Culex vectors of St. Louis encephalitis virus based on interspecies sequence variation in ribosomal DNA spacers. Am J Trop Med Hyg 53 :105–109.

    • Search Google Scholar
    • Export Citation
  • 9

    Debrunner-Vossbrinck BA, Vosbrinck CR, Vodkin MH, Novak RJ, 1996. Restriction analysis of the ribosomal DNA internal transcript spacer region of Culex restuans mosquitoes in the Culex pipiens complex. J Am Mosq Control Assoc 13 :477–482.

    • Search Google Scholar
    • Export Citation
  • 10

    Aspen S, Savage HM, 2003. Polymerase chain reaction assay identifies North American members of the Culex pipiens complex based on nucleotide sequence differences in the acetyl-cholinesterase gene Ace.2. J Am Mosq Control Assoc 19 :323–328.

    • Search Google Scholar
    • Export Citation
  • 11

    Bourguet D, Fonseca D, Vourch G, Dubois MP, Chandre F, Severini C, Raymond M, 1998. The acetylcholinesterase gene Ace: a diagnostic marker for the Pipiens and Quinquefasciatus forms of the Culex pipiens complex. J Am Mosq Control Assoc 14 :390–396.

    • Search Google Scholar
    • Export Citation
  • 12

    Smith JL, Fonseca DM, 2004. Rapid assays for identification of members of the Culex (Culex) pipiens complex, their hybrids, and other sibling species (Diptera: culicidae). Am J Trop Med Hyg 70 :339–345.

    • Search Google Scholar
    • Export Citation
  • 13

    Fonseca DM, Smith JL, Wilkerson RC, Fleischer RC, 2006. Pathways of expansion and multiple introductions illustrated by large genetic differentiation among worldwide populations of the southern house mosquito. Am J Trop Med Hyg 74 :284–289.

    • Search Google Scholar
    • Export Citation
  • 14

    Darsie RF Jr, Ward RA, 1989. Review of new Nearctic mosquito distributional records north of Mexico, with notes on additions and taxonomic changes of the fauna, 1982–89. J Am Mosq Control Assoc 5 :552–557.

    • Search Google Scholar
    • Export Citation
  • 15

    Apperson CS, Harrison BA, Unnasch TR, Hassan HK, Irby WS, Savage HM, Aspen SE, Watson DW, Rueda LM, Engber BR, Nasci RS, 2002. Host-feeding habits of Culex and other mosquitoes (Diptera: Culicidae) in the Borough of Queens in New York City, with characters and techniques for identification of Culex mosquitoes. J Med Entomol 39 :777–785.

    • Search Google Scholar
    • Export Citation
  • 16

    Barr AR, 1957. Distribution of Culex pipiens and Culex quinquefasciatus in North America. Am J Trop Med 4 :153–165.

  • 17

    Thompson JD, Higgins DG, Gibson TJ, 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22 :4673–4680.

    • Search Google Scholar
    • Export Citation
  • 18

    Lampman R, Slamecka M, Krasavin N, Kunkel K, Novak R, 2006. Culex population dynamics and West Nile transmission in east-central Illinois. J Am Mosq Control Assoc 22 :390–400.

    • Search Google Scholar
    • Export Citation
  • 19

    Gu W, Lampman R, Novak RJ, 2004. Assessment of arbovirus vector infection rates using variable size pooling. Med Vet Entomol 18 :200–204.

    • Search Google Scholar
    • Export Citation
  • 20

    Bahnck CM, Fonseca DM, 2006. Rapid assay to identify the two genetic forms of Culex (Culex) pipiens L. (Diptera: Culicidae) and hybrid populations. Am J Trop Med Hyg 75 :251–255.

    • Search Google Scholar
    • Export Citation
  • 21

    Fonseca DM, Keyghobadi N, Malcolm CA, Mehmet C, Schaffner F, Mogi M, Fleischer RC, Wilkerson RC, 2004. Emerging vectors in the Culex pipiens complex. Science 303 :1535–1538.

    • Search Google Scholar
    • Export Citation
  • 22

    Darbro JM, Harrington LC, 2006. Bird-baited traps for surveillance of West Nile mosquito vectors: effect of bird species, trap height, and mosquito escape rates. J Med Entomol 43 :83–92.

    • Search Google Scholar
    • Export Citation
  • 23

    McAvin JC, Bowles DE, Swaby JA, Blount KW, Blow JA, Quintana M, Hickman JR, Atchley DH, Niemeyer DM, 2005. Identification of Aedes aegypti and its respective life stages by real-time polymerase chain reaction. Mil Med 170 :1060–1065.

    • Search Google Scholar
    • Export Citation
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 

 

A Real-Time TaqMan Polymerase Chain Reaction for the Identification of Culex Vectors of West Nile and Saint Louis Encephalitis Viruses in North America

View More View Less
  • 1 Illinois Natural History Survey, Champaign, Illinois
Restricted access

In North America, West Nile and St. Louis encephalitis viruses have been detected in a wide range of vector species, but the majority of isolations continue to be from pools of mixed mosquitoes in the Culex subgenus Culex. Unfortunately, the morphologic identification of these important disease vectors is often difficult, particularly in regions of sympatry. We developed a sensitive real-time TaqMan polymerase chain reaction assay that allows reliable identification of Culex mosquitoes including Culex pipiens pipiens, Cx. p. quinquefasciatus, Cx. restuans, Cx. salinarius, Cx. nigripalpus, and Cx. tarsalis. Primers and fluorogenic probes specific to each species were designed based on sequences of the acetylcholinesterase gene (Ace2). Both immature and adult mosquitoes were successfully identified as individuals and as mixed species pools. This identification technique provides the basis for a rapid, sensitive, and high-throughput method for expounding the species-specific contribution of vectors to various phases of arbovirus transmission.

Save