Development and evaluation of a polymerase chain reaction (PCR) assay for the detection of Opisthorchis viverrini in fish.
Human opisthorchiais caused by the liver fluke Opisthorchis viverrini is a major fish-borne trematode infection endemic in the Southeast Asian countries. The infection is acquired through consumption of raw fish harboring metacercariae of O. viverrini. Owing to potential risk of transmission of opisthorchiasis through fish trade, rapid and reliable detection methods have gained importance to ensure food safety. In the study described here, we report successful development and evaluation of a polymerase chain reaction (PCR) assay for the detection of O. viverrini, based on the nucleotide sequence derived in this study. The assay is specific with no cross-reaction with other trematodes commonly found in fish including the closely related species, Clonorchis sinensis. The sensitivity of the assay was determined to be 10(-12)ng of O. viverrini DNA while in artificially spiked fish meat, 3 metacercariae could be detected. The results suggest that the PCR method described here is specific to O. viverrini with potential application in fish quarantining.
Parvathi
Ammini
A
Department of Fishery Microbiology, Karnataka Veterinary, Animal and Fisheries Sciences University, College of Fisheries, Mangalore 575 002, India.
Umesha
Kanasinakatte R
KR
Kumar
Sanath
S
Sithithaworn
Paiboon
P
Karunasagar
Indrani
I
Karunasagar
Iddya
I
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005393:Fish Diseases / Q000175:diagnosis* / Q000469:parasitology*; D005399:Fishes; D008969:Molecular Sequence Data; D009889:Opisthorchiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D009891:Opisthorchis / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018509:DNA, Helminth