Rapid identification and differentiation of Fasciola hepatica and Fasciola gigantica by a loop-mediated isothermal amplification (LAMP) assay

• Published in: Veterinary parasitology 2010-12, Vol.174 (3-4), p.228-233
• Main Authors: Ai, L., Li, C., Elsheikha, H.M., Hong, S.J., Chen, J.X., Chen, S.H., Li, X., Cai, X.Q., Chen, M.X., Zhu, X.Q.
• Format: Article
• Language: English
• Subjects: Animals; diagnostic sensitivity; diagnostic specificity; disease detection; disease diagnosis; Fasciola - classification; Fasciola gigantica; Fasciola hepatica; Fascioliasis; Loop-mediated isothermal amplification (LAMP); Nucleic Acid Amplification Techniques - veterinary; pathogen identification; PCR; polymerase chain reaction; Rapid detection; Sensitivity and Specificity; species specificity
• ISSN: 0304-4017; 1873-2550
• DOI: 10.1016/j.vetpar.2010.09.005

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 45min under isothermal conditions of 61°C or 62°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 104 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.