title: Loop-mediated isothermal amplification applied to filarial parasites detection in the mosquito vectors: Dirofilaria immitis as a study model.
abstract: BACKGROUND
  Despite recent advances in our understanding of the basic biology behind transmission of zoonotic infectious diseases harbored by arthropod vectors these diseases remain threatening public health concerns. For effective control of vector and treatment, precise sampling indicating the prevalence of such diseases is essential. With an aim to develop a quick and simple method to survey zoonotic pathogen-transmitting vectors, LAMP (loop-mediated isothermal amplification) was applied to the detection of filarial parasites using a filarial parasite-transmitting experimental model that included one of the mosquito vectors, Aedes aegypti, and the canine heartworm, Dirofilaria immitis.
  
  
  RESULTS
  LAMP reactions amplifying the cytochrome oxidase subunit I gene demonstrated high sensitivity when a single purified D. immitis microfilaria was detected. Importantly, the robustness of the LAMP reaction was revealed upon identification of an infected mosquito carrying just a single parasite, a level easily overlooked using conventional microscopic analysis. Furthermore, successful detection of D. immitis in wild-caught mosquitoes demonstrated its applicability to field surveys.
  
  
  CONCLUSIONS
  Due to its simplicity, sensitivity, and reliability, LAMP is suggested as an appropriate diagnostic method for routine diagnosis of mosquito vectors carrying filarial parasites. This method can be applied to the survey of not only canine filariasis but also lymphatic filariasis, another major public health problem. Therefore, this method offers great promise as a useful diagnostic method for filarial parasite detection in endemic filariasis regions.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. fukumoto@obihiro.ac.jp.
authors.forename: Hiroka
authors.initials: H
authors.lastname: Aonuma
authors.forename: Aya
authors.initials: A
authors.lastname: Yoshimura
authors.forename: Namal
authors.initials: N
authors.lastname: Perera
authors.forename: Naoaki
authors.initials: N
authors.lastname: Shinzawa
authors.forename: Hironori
authors.initials: H
authors.lastname: Bando
authors.forename: Sugao
authors.initials: S
authors.lastname: Oshiro
authors.forename: Bryce
authors.initials: B
authors.lastname: Nelson
authors.forename: Shinya
authors.initials: S
authors.lastname: Fukumoto
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Kanuka