Identification of novel Glutathione S-Transferases epsilon 2 mutation in Anopheles maculipennis s.s. (Diptera: Culicidae)

Anopheles maculipennis complex comprises some important malaria vectors in Iran, Middle East, and Europ. The
principal way to control of malaria remains on the use of chemical insecticides against its vectors because there is
no vaccine for malaria prevention. Extensive use of organophosphate compounds has caused to emergence and
distribution of insecticide resistance in Anopheles species in Asia. The current study aimed to the detection of three
well-known amino acid substitutions (I114T, L119F, and F120L) in the Glutathione S-Transferases epsilon 2
(GSTe2) gene are associated with DDT and organophosphate insecticides resistance in an Anopheles maculipennis
population collected from Iran. Adult samples of An. maculipennis were collected by hand and Total catch in
Animal and Human Shelters from Azerbaijan-Gharbi and Zanjan provinces. Following morphological identifica-
tion, DNA was extracted by YTA Genomic DNA Extraction Mini Kit for amplification of rDNA-ITS2 and GSTe2
fragments. ~500 bp fragment was amplified using F rDNA-ITS2 and GSTe2 primers. rDNA-ITS2 sequence analysis
showed 100% similarity with An. maculipennis. GSTe2 nucleotide sequence similarity within species was
99
–
100%, while, it was 95
–
96 % when compared with Anopheles sacharovi GSTe2 sequences available in Gen-
Bank. Amino acid sequence comparisons showed a novel amino acid substitution in N148D position with 15.79%
frequency. The current study reports new GSTe2 amino acid substitution in An. maculipennis s.s., for the first time.
The function of the mutation N148D and its association with resistance phenotype need to validate. However, the
integration of these data into the malaria control program still remains a challenge.