Zhu Guoding, Zhou Huayun, Li Julin, Tang Jianxia, Bai Liang, Wang Weiming, Gu Yaping, Liu Yaobao, Lu Feng, Cao Yuanyuan, Zhang Chao, Xu Sui, Cao Jun, Gao Qi
Key Laboratory of Parasitic Disease Control and Prevention (Ministry of Health), and Jiangsu Provincial Key Laboratory of Parasite Molecular Biology, Jiangsu Institute of Parasitic Diseases, Wuxi, Jiangsu Province, People's Republic of China.
Department of Parasitology, Medical College of Soochow University, Suzhou, 215123, People's Republic of China.
Parasit Vectors. 2014 Dec 11;7:582. doi: 10.1186/s13071-014-0582-7.
Anopheles sinensis is one of the most important malaria vectors in Asian countries. The rapid spread of insecticide resistance has become a major obstacle for insecticide-based strategies for vector control. Therefore, it is necessary to prepare an insecticide-resistant strain of An. sinensis to further understand the insecticide resistance mechanisms in this species to facilitate genetic approaches to targeting the insecticide-resistant population of this important malaria vector.
An. sinensis mosquitoes were collected from regions where pyrethroid resistance had been reported. The mosquitoes were subjected to continuous pyrethroid selection after species confirmation, and the forced copulation method was used to increase the mating rate. In addition, the knockdown-resistance (kdr) mutation frequencies of each generation of An. sinensis were measured; and the metabolic enzyme activities of cytochrome P450 monoxygenases (P450s) and glutathione S-transferases (GSTs) were detected.
The identification of field-captured An. sinensis was confirmed by both morphological and molecular methods. The population of An. sinensis exhibited stable resistance to pyrethroid after continuous generations of pyrethroid selection in the laboratory with high kdr mutation frequencies; and elevated levels of both P450s and GSTs were significantly found in field selected populations comparing with the laboratory susceptible strain. So far, the colonised strain has reached its eleventh generation and culturing well in the laboratory.
We colonised a pyrethroid-resistant population of An. sinensis in the laboratory, which provides a fundamental model for genetic studies of this important malaria vector.
中华按蚊是亚洲国家最重要的疟疾传播媒介之一。杀虫剂抗性的迅速传播已成为基于杀虫剂的媒介控制策略的主要障碍。因此，有必要培育中华按蚊的抗杀虫剂品系，以进一步了解该物种的抗杀虫剂机制，从而促进针对这一重要疟疾媒介抗杀虫剂种群的遗传方法研究。
从报告有拟除虫菊酯抗性的地区采集中华按蚊。确认物种后，对这些蚊子进行连续的拟除虫菊酯筛选，并采用强制交配法提高交配率。此外，测定了每一代中华按蚊的击倒抗性（kdr）突变频率；并检测了细胞色素P450单加氧酶（P450s）和谷胱甘肽S-转移酶（GSTs）的代谢酶活性。
通过形态学和分子方法确认了野外捕获的中华按蚊。在实验室连续多代进行拟除虫菊酯筛选后，中华按蚊种群对拟除虫菊酯表现出稳定的抗性，kdr突变频率较高；与实验室敏感品系相比，在野外筛选种群中显著发现P450s和GSTs水平均升高。到目前为止，定殖品系已传至第十一代，在实验室中培养良好。
我们在实验室定殖了一个对拟除虫菊酯具有抗性的中华按蚊种群，这为对这一重要疟疾媒介进行遗传研究提供了一个基础模型。
