Applied Parasitology Research Laboratory, Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok, 10110, Thailand.
Applied Parasitology Research Laboratory, Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok, 10110, Thailand; Center of Excellence in Animal, Plant and Parasitic Biotechnology, Srinakharinwirot University, Bangkok, 10110, Thailand.
Vet Parasitol. 2021 Apr;292:109396. doi: 10.1016/j.vetpar.2021.109396. Epub 2021 Mar 1.
Cestodes belonging to the genus Raillietina are a major veterinary health problem in the poultry industry, especially in chickens (Gallus gallus domesticus) and ducks (Anas playtrhynchos domesticus). In this study, loop-mediated isothermal amplification coupled with a lateral flow dipstick (LAMP-LFD) assay was established and validated for the detection of R. echinobothrida, R. tetragona, and R. cesticillus in one reaction. The LAMP-LFD assay can be completed in 75 min under isothermal conditions at 66 °C and the results can be obtained by observation with the naked eye. This assay was very specific and had no cross-amplification with other closely related parasites (Cotugnia sp., Diorchis formosensis, Fimbriaria sp., Echinostoma sp., E. miyagawai, Hypoderaeum conoideum, Prosthogonimus cuneatus, and Ascaridia galli) or their definitive hosts (G. g. domesticus, A. p. domesticus). The sensitivity of the LAMP-LFD assay was detected with three Raillietina species at 0.5 ng, which was enough for gravid proglottid DNA detection. The accuracy test showed that the LAMP-LFD assay demonstrated accurate verification results when compared to morphological results. This is a novel LAMP-LFD assay that is highly specific and sensitive for the detection of Raillietina species. It can be applied to detection for epidemiological investigations, monitoring programs, surveillance, control, and to solve veterinary health problems for the poultry industry in Raillietina endemic areas.
绦虫属的绦虫是家禽业的主要兽医健康问题，特别是在鸡（Gallus gallus domesticus）和鸭（Anas playtrhynchos domesticus）中。在这项研究中，建立并验证了一种环介导等温扩增结合侧流层析（LAMP-LFD）检测方法，用于在一个反应中检测 R. echinobothrida、R. tetragona 和 R. cesticillus。LAMP-LFD 检测法可以在 66°C 的等温条件下在 75 分钟内完成，并且可以通过肉眼观察结果。该检测方法非常特异，与其他密切相关的寄生虫（Cotugnia sp.、Diorchis formosensis、Fimbriaria sp.、Echinostoma sp.、E. miyagawai、Hypoderaeum conoideum、Prosthogonimus cuneatus 和 Ascaridia galli）或其终末宿主（G. g. domesticus、A. p. domesticus）没有交叉扩增。通过三种绦虫检测 LAMP-LFD 检测法的灵敏度为 0.5ng，足以检测到有孕节片的 DNA。准确性测试表明，与形态学结果相比，LAMP-LFD 检测法的验证结果准确。这是一种新型的 LAMP-LFD 检测方法，对绦虫属的检测具有高度的特异性和敏感性。它可以应用于流行病学调查、监测计划、监测、控制，以及解决绦虫流行地区家禽业的兽医健康问题。
