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

被引：50
作者：
Ai, L. [2 ,5 ]
Li, C. [2 ]
Elsheikha, H. M. [3 ]
Hong, S. J. [4 ]
Chen, J. X. [5 ]
Chen, S. H. [5 ]
Li, X. [2 ]
Cai, X. Q. [6 ]
Chen, M. X. [5 ]
Zhu, X. Q. [1 ,7 ]
机构：
[1] CAAS, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol,Dept Parasitol, Lanzhou 730046, Gansu, Peoples R China
[2] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China
[3] Univ Nottingham, Sch Vet Med & Sci, Loughborough LE12 5RD, Leics, England
[4] Chung Ang Univ, Coll Med, Dept Med Environm Biol, Seoul 156756, South Korea
[5] Chinese Ctr Dis Control & Prevent, Natl Inst Parasit Dis, Shanghai 200025, Peoples R China
[6] Zhongshan Entry Exit Inspect & Quarantine Bur, Zhongshan 528403, Guangdong, Peoples R China
[7] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan Province, Peoples R China
来源：
VETERINARY PARASITOLOGY | 2010年 / 174卷 / 3-4期
关键词：
Fasciola hepatica; Fasciola gigantica; Fascioliasis; Loop-mediated isothermal amplification (LAMP); Rapid detection; PCR; MOLECULAR CHARACTERIZATION; LYMNAEA-VIATRIX; TRIPLOID FORMS; RIBOSOMAL DNA; F-HEPATICA; PCR; SEQUENCES; SAMPLES;
D O I：
10.1016/j.vetpar.2010.09.005
中图分类号：
R38 [医学寄生虫学]; Q [生物科学];
学科分类号：
07 ; 0710 ; 09 ; 100103 ;
摘要：
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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页码：228 / 233
页数：6
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