Open AccessDevelopment of a Polymerase Chain Reaction Assay for Differentiation Between Culex pipiens pipiens and Cx. p. quinquefasciatus (Diptera: Culicidae) in North America Based on Genomic Differences Identified by Subtractive Hybridization
Author(s) -
Mary B. Crabtree,
Harry M. Savage,
Barry R. Miller
Publication year - 1997
Publication title -
journal of medical entomology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.866
H-Index - 99
eISSN - 1938-2928
pISSN - 0022-2585
DOI - 10.1093/jmedent/34.5.532
Subject(s) - biology , suppression subtractive hybridization , polymerase chain reaction , culex quinquefasciatus , genomic dna , culex pipiens , dna , dna sequencing , genome , genetics , microbiology and biotechnology , virology , complementary dna , gene , cdna library , botany , aedes aegypti , larva
Culex pipiens is a complex of mosquitoes that are involved in the transmission of pathogens, including St. Louis encephalitis virus in North America. The 2 major taxa in the complex, Cx. p. pipiens and Cx. p. quinquefasciatus, are nearly identical morphologically, making identification of field-collected specimens difficult, and attempts at differentiation based on biochemical and molecular techniques have been unsuccessful. We report here the use of genomic subtractive hybridization to identify a region of nucleic acid heterology between the genomes of Cx. p. pipiens and Cx. p. quinquefasciatus and the development of a polymerase chain reaction (PCR) assay to discriminate between them. PCR primers based on the nucleic acid sequence of a Cx. p. pipiens-unique DNA fragment were used to differentiate Cx. p. pipiens and Cx. p. pipiens/quinquefasciatus hybrids from Cx. p. quinquefasciatus by using extracted individual mosquito genomic DNA, crude DNA preparations from a mosquito head or legs, and DNA from triturated mosquito pools.