Detection of the Plasmodium falciparum Kelch-13 gene P553L mutation in sporozoites isolated from mosquito salivary glands in South-Central Vietnam.
Detection of the Plasmodium falciparum Kelch-13 gene P553L mutation in sporozoites isolated from mosquito salivary glands in South-Central Vietnam.
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DOI：
10.1186/s13071-017-2247-9
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发表时间：
2017-06-24
影响因子：
3.2
通讯作者：
Marchand RP
中科院分区：
文献类型：
--
作者：
Maeno Y;Quang NT;Culleton R;Kawai S;Masuda G;Hori K;Nakazawa S;Marchand RP
在东南亚，恶性疟原虫已经对青蒿素产生了抗药性。恶性疟原虫Kelch-13（Pfk 13）基因突变与体外和体内青蒿素耐药性相关。我们调查了从大劣按蚊唾液腺分离的子孢子阶段寄生虫PfK 13突变的患病率。在越南中南部的庆富公社内的两个地点，蚊子被人类登陆捕获。根据形态学特征和核苷酸序列分析进行按蚊种的鉴定。将子孢子感染的唾液腺储存在滤纸上并在4-6 °C下。一个巢式PCR靶向小亚基核糖体RNA基因用于疟原虫种鉴定。通过巢式PCR扩增Pfk 13，并进行核苷酸测序。33个恶性疟原虫子孢子样品中有5个在PfK 13位点携带P553 L突变。这种突变以前在越南有记录，但在庆和省没有，因为以前没有进行过K13多态性的调查。这些结果表明，蚊子阶段疟疾寄生虫样品的抗药性的分子流行病学研究的效用。
Plasmodium falciparum has developed resistance against artemisinin in Southeast Asia. Mutations in the P. falciparum Kelch-13 (Pfk13) gene are associated with artemisinin resistance in vitro and in vivo. We investigated the prevalence of mutations in PfK13 from sporozoite-stage parasites isolated from the salivary glands of Anopheles dirus mosquitoes. Mosquitoes were caught by human-landing catches at two locations within the Khanh Phu commune, South-Central Vietnam. Identification of Anopheles species was performed based on morphological features and nucleotide sequence analysis. Sporozoite-infected salivary glands were stored on filter paper and at 4–6 °C. A nested-PCR targeting the small subunit ribosomal RNA gene was used for Plasmodium species identification. Pfk13 was amplified by nested PCR, and subjected to nucleotide sequencing. Five of 33 P. falciparum sporozoite samples carried the P553L mutation at the PfK13 locus. This mutation has been recorded previously in Vietnam, but not in Khanh Hoa province, were surveys of K13 polymorphism have not previously been carried out. These results demonstrate the utility of mosquito-stage malaria parasite samples for studies on the molecular epidemiology of drug resistance.