Clonorchis sinensis: development and evaluation of a nested polymerase chain reaction (PCR) assay.
Clonorchis sinensis is a fish-borne trematode endemic to East Asia, which infects over 35 million people globally. In the study described here, we developed a nested polymerase chain reaction (PCR) method for the specific and reliable detection of C. sinensis. The primers designed from the nucleotide sequence data derived in this study were evaluated for their specificity and sensitivity for the detection of C. sinensis. The specific amplification products were obtained only with C. sinensis and no amplifications occurred with the DNA of closely related trematodes including Opisthorchis viverrini demonstrating the specificity of the assay. The novel PCR method described here will be useful for the quarantine of fishery products and evaluation of transmission status of clonorchiasis in the endemic areas.
Parvathi
Ammini
A
Department of Fishery Microbiology, Karnataka Veterinary, Animal and Fisheries Sciences University, College of Fisheries, Mangalore, India.
Sanath Kumar
H
H
Kenchanna Prakasha
B
B
Lu
Jieyuan
J
Xu
Xuenian
X
Hu
Wei
W
Feng
Zheng
Z
Karunasagar
Indrani
I
Karunasagar
Iddya
I
D000818:Animals; D001483:Base Sequence; D003003:Clonorchiasis / Q000175:diagnosis / Q000517:prevention & control / Q000635:transmission; D003004:Clonorchis sinensis / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000737:chemistry / Q000592:standards; D018509:DNA, Helminth / Q000737:chemistry; D005393:Fish Diseases / Q000175:diagnosis / Q000469:parasitology / Q000635:transmission; D005399:Fishes; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000592:standards*; D019105:Random Amplified Polymorphic DNA Technique; D017747:Seafood / Q000469:parasitology; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018509:DNA, Helminth