Ultrasensitive malaria detection system for Anopheles mosquito field surveillance using droplet digital PCR

Tamasa Araki; Akihide Koyama; Hiro Yoshimura; Ayako Arai; Satoru Kawai; Shuto Sekizawa; Yuko Umeki; Yumiko Saito-Nakano; Takashi Imai; Munehiro Okamoto; Megumi Sato; Wipaporn Thabthimthong; Taratorn Kemthong; Hajime Hisaeda; Suchinda Malaivijitnond; Takeshi Annoura

doi:10.1016/j.parint.2024.102891

Ultrasensitive malaria detection system for Anopheles mosquito field surveillance using droplet digital PCR

Parasitol Int. 2024 Mar 26:101:102891. doi: 10.1016/j.parint.2024.102891. Online ahead of print.

Authors

Tamasa Araki¹, Akihide Koyama², Hiro Yoshimura³, Ayako Arai¹, Satoru Kawai⁴, Shuto Sekizawa³, Yuko Umeki⁵, Yumiko Saito-Nakano⁵, Takashi Imai⁵, Munehiro Okamoto⁶, Megumi Sato⁷, Wipaporn Thabthimthong⁸, Taratorn Kemthong⁸, Hajime Hisaeda¹, Suchinda Malaivijitnond⁹, Takeshi Annoura¹⁰

Affiliations

¹ Department of Parasitology, National Institute of Infectious Diseases, Toyama, Shinjuku-ku, Tokyo, Japan.
² Department of Legal Medicine, Graduate School of Medical and Dental Science, Niigata University, Asahimachi, Chuo-ku, Niigata, Japan.
³ Department of Parasitology, National Institute of Infectious Diseases, Toyama, Shinjuku-ku, Tokyo, Japan; Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo, Japan.
⁴ Department of Tropical Medicine and Parasitology, Dokkyo Medical University, Mibu, Tochigi, Japan.
⁵ Department of Parasitology, National Institute of Infectious Diseases, Toyama, Shinjuku-ku, Tokyo, Japan; Antimicrobial Resistance Research Center, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan.
⁶ Section of Molecular Biology, Center for the Evolutionary Origins of Human Behavior, Kyoto University, Inuyama, Aichi, Japan.
⁷ Graduate School of Health Sciences, Niigata University, Niigata, Japan.
⁸ National Primate Research Center of Thailand, Chulalongkorn University, Saraburi, Thailand.
⁹ National Primate Research Center of Thailand, Chulalongkorn University, Saraburi, Thailand; Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
¹⁰ Department of Parasitology, National Institute of Infectious Diseases, Toyama, Shinjuku-ku, Tokyo, Japan; Cooperative Division of Veterinary Sciences, Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Tokyo, Japan. Electronic address: annoura@niid.go.jp.

PMID: 38537686
DOI: 10.1016/j.parint.2024.102891

Abstract

Malaria remains a significant global public health concern, with a recent increase in the number of zoonotic malaria cases in Southeast Asian countries. However, limited reports on the vector for zoonotic malaria exist owing to difficulties in detecting parasite DNA in Anopheles mosquito vectors. Herein, we demonstrate for the first time that several Anopheles mosquitoes contain simian malaria parasite DNA using droplet digital PCR (ddPCR), a highly sensitive PCR method. An entomological survey was conducted to identify simian malaria vector species at Phra Phothisat Temple (PPT), central Thailand, recognized for a high prevalence of simian malaria in wild cynomolgus macaques. A total of 152 mosquitoes from six anopheline species were collected and first analyzed by a standard 18S rRNA nested-PCR analysis for malaria parasite which yielded negative results in all collected mosquitoes. Later, ddPCR was used and could detect simian malaria parasite DNA, i.e. Plasmodium cynomolgi, in 25 collected mosquitoes. And this is the first report of simian malaria parasite DNA detection in Anopheles sawadwongporni. This finding proves that ddPCR is a powerful tool for detecting simian malarial parasite DNA in Anopheles mosquitoes and can expand our understanding of the zoonotic potential of malaria transmission between monkeys and humans.

Keywords: Anopheles; Droplet digital PCR; Entomological field survey; Plasmodium cynomolgi; Simian malaria; Zoonotic malaria.