Rapid identification of Plasmodium-carrying mosquitoes using loop-mediated isothermal amplification |
| |
Authors: | Aonuma Hiroka Suzuki Moemi Iseki Hiroshi Perera Namal Nelson Bryce Igarashi Ikuo Yagi Takeshi Kanuka Hirotaka Fukumoto Shinya |
| |
Institution: | a National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan b KOKORO Biology Group, Laboratories for Integrated Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan |
| |
Abstract: | With an aim to develop a quick and simple method to survey pathogen-transmitting vectors, LAMP (loop-mediated isothermal amplification) was applied to the identification of Plasmodium-carrying mosquitoes, specifically a Plasmodium-transmitting experimental model using rodent malaria parasite (Plasmodium berghei) and anopheline mosquitoes (Anopheles stephensi). The detection sensitivity limit of the LAMP reaction amplifying the SPECT2 gene was determined to be 1 × 102 purified Plasmodium parasites, estimated to be sufficient for reliable identification of infectious mosquitoes. The robustness of the LAMP reaction was revealed by its ability to detect both Plasmodium oocysts and sporozoites from an “all-in-one” template using whole mosquito bodies. Moreover, LAMP successfully identified an infectious mosquito carrying just a single oocyst in its midgut, a level that can be easily overlooked in conventional microscopic analysis. These observations suggest that LAMP is more reliable and useful for routine diagnosis of vector mosquitoes in regions where vector-borne diseases such as malaria are endemic. |
| |
Keywords: | LAMP Diagnosis Plasmodium Mosquito Malaria |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|