Figure 1.
A map of China showing the distribution of mosquito sampling sites.
Site name: 1, Mengla county (Yunnan Province); 2, Yuanyang county (Yunnan Province); 3, Liuyang county (Hunan Province); 4, Wuxue county (Hubei Province); and 5, Sihong city (Jiangsu Province).
Table 1.
Primer sequence of multiplex allele-specific PCR for the detection of mutations in codon 1014 of para-type sodium channel gene and super kdr mutation in codon 918 in Anopheles sinensis.
Figure 2.
Mortality rates of Anopheles sinensis in standard WHO tube bioassays.
A: after exposure to 0.05% deltamethrin test papers and 24 hr recovery period. B: pairwise Tukey–Kramer HSD test for statistical significance in mortality rates. The vertical bar stands for standard error of the mean.
Figure 3.
Examples of nucleotide sequence chromatograms of kdr genotypes detected in Anopheles sinensis from China.
The position at codon 1014 of the para-type sodium channel gene is indicated by a rectangle box. A: four types of homozygote genotypes detected; and B: six types of heterozygote genotypes detected (K = G/T; Y = T/C; S = G/C).
Figure 4.
Allele-specific PCR (AS-PCR) for kdr genotyping in Anopheles sinensis.
A: Schematic representation of the primer location and predicted size of the PCR product. Arrow indicated the location of PCR primers in which sequences are reported in Table 1. Primer pair TTG-F and kdr-R amplifies a 141 bp fragment for the wildtype allele (for codon TTG). Primer pair TGT-F/kdr-R yields a 158 bp fragment for L1014C allele (codon TTC). Similarly, primer pair kdr-F/TTT-R and kdr-F/TTC-R leads to amplification of a 221 bp and 241 bp fragment diagnostic to the L1014F allele (for codon TTT and TTC), respectively. B: An example of AS-PCR gel electrophoresis. Lane M: 100 bp DNA ladder; lanes 1–4: homozygote for the codons TTG, TGT, TTT, and TTC, respectively; lanes 5–10: six heterozygous individuals (TTG/TTT, TGT/TTT, TTG/TGT, TTT/TTC, TGT/TTC, and TTG/TTC).
Table 2.
Frequencies (in percentage) of kdr alleles in relation to mosquito survival phenotype determined by the deltamethrin susceptibility bioassay in five Anopheles sinensis populations from China.
Table 3.
Glutathione s-transferases (GSTs) and monooxygenases activity in Anopheles sinensis female adults in five populations in China when compared with lab susceptible strain.
Figure 5.
Relationship between kdr, metabolic detoxifying enzyme activity and pyrethroid resistance in Anopheles sinensis.
Association between kdr genotypes and survival rate of mosquitoes after the standard WHO tube deltamethrin susceptibility bioassay (A), and variation of resistant (survived the bioassay) and susceptible (died in the bioassay) individuals in glutathione s-transferases activity (B) and monooxygenases activity (C). **, P < 0.01.
Table 4.
Summary results of stepwise multivariate regression analysis on knockdown time in Anopheles sinensis.
Table 5.
Stepwise logistic regression for factors significantly associated with deltamethrin resistance in Anopheles sinensis.