title: Development and evaluation of loop-mediated isothermal amplification (LAMP) for rapid detection of Clonorchis sinensis from its first intermediate hosts, freshwater snails.
abstract: Clonorchiasis, caused by Clonorchis sinensis, is a key foodborne zoonosis, which is mainly found in China, Korea and Vietnam. Detection of this parasite from the second intermediate host, the freshwater fish is the common method for epidemiological surveys of this parasite, but is time consuming, labour intensive and easily leads to misdiagnosis. In this study, we have developed a rapid, sensitive and reliable molecular method for the diagnosis of C. sinensis from its first intermediate hosts, freshwater snails, based on a loop-mediated isothermal amplification (LAMP) method. The specific amplified fragment from genomic DNA of C. sinensis did not cross-react with those from other relevant trematodes and a range of hosts (freshwater fish, shrimps and snails) of C. sinensis living in similar environments. The detection limit of the LAMP method was as low as 10 fg which was 1000 times more sensitive than conventional PCR, which was also demonstrated by successful application to field samples. These results show that the LAMP method is a more sensitive tool than conventional PCR for the detection of C. sinensis infection in the first intermediate hosts and, due to a simpler protocol, is an ideal molecular method for field-based epidemiological surveys of this parasite.
authors.affiliation: Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering Sun Yat-Sen University, Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China.
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
authors.forename: T
authors.initials: T
authors.lastname: Wen
authors.forename: D-H
authors.initials: DH
authors.lastname: Lai
authors.forename: Y-Z
authors.initials: YZ
authors.lastname: Wen
authors.forename: Z-D
authors.initials: ZD
authors.lastname: Wu
authors.forename: T-B
authors.initials: TB
authors.lastname: Yang
authors.forename: X-B
authors.initials: XB
authors.lastname: Yu
authors.forename: G
authors.initials: G
authors.lastname: Hide
authors.forename: Z-R
authors.initials: ZR
authors.lastname: Lun
mesh_terms: D000818:Animals; D002681:China / Q000453:epidemiology; D003003:Clonorchiasis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D003004:Clonorchis sinensis / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005393:Fish Diseases / Q000453:epidemiology / Q000469:parasitology*; D005399:Fishes; D005618:Fresh Water; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D012908:Snails / Q000469:parasitology*; D015047:Zoonoses
chemical_list: D017931:DNA Primers; D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer