Abstract
BackgroundDespite 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.ResultsLAMP 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.ConclusionDue 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.
Highlights
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
Optimization of Loop-mediated isothermal amplification (LAMP) reaction conditions using the described primer set revealed ideal settings to be 63°C and 70 minutes. Sequencing of these LAMP products showed that amplified DNA were of D. immitis cytochrome oxidase subunit I gene
D. immitis microfilariae DNA obtained from cultured adult worms was used as a template for LAMP reactions and yielded an amplified product approximately 30 minutes after incubation using DNA from 1 × 104 parasites (152 ng) (Fig. 2A)
Summary
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. Human clinical cases of diseases, such as dirofilariasis, babesiosis, and leishmaniases, all caused by parasites transmitted by arthropod vectors from animals, have been reported to be on the rise [2,3,4,5,6]. Infection with these diseases have affects on humans and on domestic and wild animals that can serve as potential reservoirs by hosting pathogens long-term despite being asymptomatic. Control of mosquitoes based on precise surveillance of Dirofilaria species is essential for preventing infection of both domestic animals and humans with these parasites
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
