Detection of the Plasmodium falciparum Kelch-13 gene P553L mutation in sporozoites isolated from mosquito salivary glands in South-Central Vietnam

Yoshimasa Maeno, Nguyen Tuyen Quang, Richard Culleton, Satoru Kawai, Gaku Masuda, Kaoru Hori, Shusuke Nakazawa, Ron P. Marchand

Research output: Contribution to journalArticle

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Abstract

Background: 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. Methods: 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. Results: 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. Conclusion: These results demonstrate the utility of mosquito-stage malaria parasite samples for studies on the molecular epidemiology of drug resistance.

Original languageEnglish
Article number308
JournalParasites and Vectors
Volume10
Issue number1
DOIs
Publication statusPublished - 24-06-2017

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Sporozoites
Vietnam
Plasmodium falciparum
Salivary Glands
Culicidae
Mutation
Anopheles
Genes
Parasites
Polymerase Chain Reaction
Southeastern Asia
Plasmodium
Molecular Epidemiology
rRNA Genes
Drug Resistance
Malaria
Sequence Analysis
Nucleotides
artemisinine

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Infectious Diseases

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Maeno, Yoshimasa ; Quang, Nguyen Tuyen ; Culleton, Richard ; Kawai, Satoru ; Masuda, Gaku ; Hori, Kaoru ; Nakazawa, Shusuke ; Marchand, Ron P. / Detection of the Plasmodium falciparum Kelch-13 gene P553L mutation in sporozoites isolated from mosquito salivary glands in South-Central Vietnam. In: Parasites and Vectors. 2017 ; Vol. 10, No. 1.
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Detection of the Plasmodium falciparum Kelch-13 gene P553L mutation in sporozoites isolated from mosquito salivary glands in South-Central Vietnam. / Maeno, Yoshimasa; Quang, Nguyen Tuyen; Culleton, Richard; Kawai, Satoru; Masuda, Gaku; Hori, Kaoru; Nakazawa, Shusuke; Marchand, Ron P.

In: Parasites and Vectors, Vol. 10, No. 1, 308, 24.06.2017.

Research output: Contribution to journalArticle

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AU - Maeno, Yoshimasa

AU - Quang, Nguyen Tuyen

AU - Culleton, Richard

AU - Kawai, Satoru

AU - Masuda, Gaku

AU - Hori, Kaoru

AU - Nakazawa, Shusuke

AU - Marchand, Ron P.

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Y1 - 2017/6/24

N2 - Background: 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. Methods: 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. Results: 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. Conclusion: These results demonstrate the utility of mosquito-stage malaria parasite samples for studies on the molecular epidemiology of drug resistance.

AB - Background: 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. Methods: 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. Results: 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. Conclusion: These results demonstrate the utility of mosquito-stage malaria parasite samples for studies on the molecular epidemiology of drug resistance.

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