polymerase chain reaction based method for the detection of Culex quinquefasciatus (Diptera: Culicidae)

Culex quinquefasciatus Say is the major vector of the filarial parasite Wuchereria bancrofti (Cobbold) which causes lymphatic filariasis in humans. A repetitive DNA sequence from the genome of C. quinquefasciatus has been cloned and completely sequenced. The 693 bp cloned fragment had an A+T content...

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Bibliographic Details
Published in:Bulletin of entomological research 2000-02, Vol.90 (1), p.63-68
Main Authors: Hettiaratchi, U.P.K, Munasingha, D.H.N, Chandrasekharan, N.V, Karunanayake, E.H, Jayasekera, N
Format: Article
Language:English
Subjects:
Animals
Anopheles vagus
Base Sequence
Biological and medical sciences
Blotting, Southern
Brackish
Culex - classification
Culex - genetics
Culex gelidus
Culex pseudovishnui
Culex quinquefasciatus
Culex tritaeniorhynchus
Culicidae
detection
disease vectors
DNA probes
Female
Freshwater
Fundamental and applied biological sciences. Psychology
genbank/af089002
Genes, Insect
Humans
identification
lymphatic filariasis
Mansonia uniformis
Medically important nuisances and vectors, pests of stored products and materials: population survey and control
Molecular Sequence Data
nucleic acid hybridization
nucleotide sequences
polymerase chain reaction
Polymerase Chain Reaction - methods
repetitive sequences
Sequence Analysis, DNA
Vectors. Intermediate hosts
Wuchereria bancrofti
Citations: Items that this one cites
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Description
Summary:Culex quinquefasciatus Say is the major vector of the filarial parasite Wuchereria bancrofti (Cobbold) which causes lymphatic filariasis in humans. A repetitive DNA sequence from the genome of C. quinquefasciatus has been cloned and completely sequenced. The 693 bp cloned fragment had an A+T content of 72%. Dot matrix analysis of the fragment did not reveal any direct or inverted repeats within it. Southern blot analysis using a variety of restriction enzymes appeared to indicate that the cloned fragment was interspersed within the genome with a copy number of approximately 30,000. A search of the GenBank database did not reveal significant homologies to any previously cloned sequences. Although the probe was sensitive enough to detect picogram quantities of DNA, it was not specific for C. quinquefasciatus, as it hybridized with DNA from other mosquito species, Culex pseudovishnui Colless, Culex gelidus Theobald, Culex tritaeniorhynchus Giles, Anopheles vagus Dönitz and Mansonia uniformis (Theobald). However PCR primers derived from the cloned sequence, IpC, were found to be specific and amplified only C. quinquefasciatus DNA. The optimized PCR assay was found to be very sensitive and was capable of detecting DNA from all stages of C. quinquefasciatus thus making it an ideal diagnostic tool.
ISSN:0007-4853
1475-2670
DOI:10.1017/s0007485300000729