Journal
VETERINARY PARASITOLOGY
Volume 174, Issue 3-4, Pages 228-233Publisher
ELSEVIER
DOI: 10.1016/j.vetpar.2010.09.005
Keywords
Fasciola hepatica; Fasciola gigantica; Fascioliasis; Loop-mediated isothermal amplification (LAMP); Rapid detection; PCR
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Funding
- Program for Changjiang Scholars and Innovative Research Team in University [IRT0723]
- State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences
- Yunnan Provincial Program for Introducing High-level Scientists [2009CI125]
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The present study developed and validated a species-specific loop-mediated isothermal amplification (LAMP) assay for the rapid detection and discrimination of Fasciola hepatica and Fasciola gigantica. The LAMP assay is inexpensive, easy to perform and shows rapid reaction, wherein the amplification can be obtained in 45 min under isothermal conditions of 61 degrees C or 62 degrees C by employing a set of four species-specific primer mixtures and results can be checked through naked-eye visualization. The optimal assay conditions with no cross-reaction with other closely related trematodes (Clonorchis sinensis, Opisthorchis viverrini, Orientobilharzia turkestanicum and Schistosoma japonicum) as well as within the two Fasciola species were established. The assay was validated by examining F. gigantica DNA in the intermediate host snails and in faecal samples. The results indicated that the LAMP assay is approximately 10(4) times more sensitive than the conventional specific PCR assays. These findings indicate that this Fasciola species-specific LAMP assay may have a potential clinical application for detection and differentiation of Fasciola species, especially in endemic countries. (C) 2010 Elsevier B.V. All rights reserved.
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