Identification of five species of the Anopheles dirus complex from Thailand, using allele-specific polymerase chain reaction

Summary The Anopheles dirus complex of mosquitoes contains some of the most important vectors of malaria in Southeast Asia. To distinguish five species of the complex that occur in Thailand, a method using the polymerase chain reaction (PCR) was developed. The method utilizes allele‐specific amplifi...

Full description

Saved in:
Bibliographic Details
Published in:Medical and veterinary entomology 1999-02, Vol.13 (1), p.24-32
Main Authors: Walton, C., Handley, J. M., Kuvangkadilok, C., Collins, F. H., Harbach, R. E., Baimai, V., Butlin, R. K.
Format: Article
Language:English
Subjects:
allele-specific polymerase chain reaction
Alleles
Animals
Anopheles - classification
Anopheles - genetics
Anopheles dirus
Anopheles nemophilous
Base Sequence
Culicidae
DNA Primers
Female
Freshwater
Genes, Insect
Humans
internal transcribed spacer 2
malaria vectors
Molecular Sequence Data
Polymerase Chain Reaction - methods
species identification
Thailand
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Summary The Anopheles dirus complex of mosquitoes contains some of the most important vectors of malaria in Southeast Asia. To distinguish five species of the complex that occur in Thailand, a method using the polymerase chain reaction (PCR) was developed. The method utilizes allele‐specific amplification to detect fixed differences between the species in the DNA sequence of the ribosomal DNA internal transcribed spacer 2. Primers were designed to amplify fragments of diagnostic length from the DNA of the different species. The method was tested on 179 mosquitoes of the An. dirus complex from many parts of Thailand and shown to be effective. Every specimen was unambiguously identified as species A, B, C, D or F (i.e. An. dirus s.s. species B, C, D or An. nemophilous, respectively) by the PCR method, with confirmation of 58/61 identifications from polytene chromosome characteristics. For the other three specimens (3/44 from Kanchanaburi 5 locality), there was disagreement between the PCR and chromosomal methods of species identification (probably due to errors in the chromosomal identifications). Primers can be combined in a single PCR reaction providing a rapid, sensitive and straightforward method of species identification. Only small quantities of DNA are required, leaving most of the mosquito to be used for other analyses.
ISSN:0269-283X
1365-2915
DOI:10.1046/j.1365-2915.1999.00142.x