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Use of the polymerase chain reaction to identify mosquito species of the Anopheles gambiae complex
Author(s) -
PASKEWITZ S. M.,
COLLINS F. H.
Publication year - 1990
Publication title -
medical and veterinary entomology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 82
eISSN - 1365-2915
pISSN - 0269-283X
DOI - 10.1111/j.1365-2915.1990.tb00453.x
Subject(s) - biology , primer (cosmetics) , polymerase chain reaction , microbiology and biotechnology , dna , anopheles gambiae , genetics , ribosomal dna , inverse polymerase chain reaction , intergenic region , nested polymerase chain reaction , gene , genome , phylogenetics , malaria , chemistry , organic chemistry , immunology
. A nonradiometric method has been developed for distinguishing between the sibling species Anopheles gambiae Giles and An.arabiensis Patton, two important Afrotropical vectors of malaria. DNA fragments of species diagnostic length are amplified by polymerase chain reaction (PCR) from a small amount of unknown DNA and three different PCR primers. All three PCR primers are based on ribosomal DNA (rDNA) sequences. A universal plus‐strand primer (A 0 ) is derived from a conserved region at the 3' end of the 28S rDNA coding region. Two species‐specific minus‐strand primers (Aa 05 and Ag 1 3 ) are derived from sequences in the intergenic spacers. The Ag 13 sequence is approximately 1.3 kb downstream of A 0 ; the Aa 0 5 sequence is about 0.5 kb downstream of A 0 . When mosquito DNA is amplified in the presence of all three primers, a 1.3 kb fragment is produced if An.gambiae DNA is used as template, and a 0.5 kb fragment is produced if An.arabiensis DNA is used. Amplification of DNA from An. gambiae I An.arabiensis hybrids produces both the 1.3 kb and the 0.5 kb fragments. Neither diagnostic fragment is produced when DNA from other species in the An.gambiae complex is used as template.