TABLE 8

PCR-based assays for detection and molecular characterization of food-borne trematodes c

SpeciesAmplification methodDetection method(s)TargetSample type(s)Reference
O. viverrini PCRGel electrophoresis O. viverrini-specific repetitive DNA fragmentStool 491
O. viverrini PCR a Gel electrophoresis O. viverrini-specific repetitive DNA fragmentStool 492
O. viverrini, C. sinensis Multiplex PCRGel electrophoresisMitochondrial DNAWorms, metacercariae, eggs 493
OpisthorchiidaePCRGel electrophoresisITS2Stool 494
O. viverrini PCR a Gel electrophoresis O. viverrini-specific repetitive DNA fragmentStool 495
O. viverrini Real-time PCRHydrolysis probe O. viverrini-specific repetitive DNA fragmentTumor tissue 368
O. viverrini PCRGel electrophoresis O. viverrini genomic DNA cloneStool 496
O. viverrini, C. sinensis, Haplorchis taichui PCR-RFLPGel electrophoresisITS2Stool 497
O. viverrini, H. taichui Nested PCRGel electrophoresis cox1 Stool 498
C. sinensis, O. felineus, O. viverrini Real-time PCRHydrolysis probeITS2Stool 369
O. viverrini Real-time PCRFRET probes-MCA O. viverrini-specific repetitive DNA fragmentStool 499
O. viverrini, H. taichui PCRGel electrophoresisITS1Stool 489
O. viverrini, H. taichui PCRGel electrophoresisITS2Stool
C. sinensis, O. viverrini, O. felineus MLPAGel electrophoresis, capillary sequencerITS1Worm 372
O. viverrini (and S. stercoralis)Real-time multiplex PCR b FRET probes-MCA O. viverrini-specific repetitive DNA fragmentStool 356
O. viverrini, H. taichui HAT-RAPD PCRGel electrophoresisGenomicMicroscopy-positive stool 500
O. viverrini LAMPVisual nad1 Stool, fish 374
O. viverrini LAMPReal time, visualITS1Stool 373
O. viverrini, C. sinensis Real-time multiplex PCR b FRET probes-MCA nad2 Stool 371
C. sinensis Real-time PCRHydrolysis probeITS1Stool, fish 370
F. hepatica, F. gigantica, intermediate Fasciola Sequence-related amplified polymorphismGel electrophoresisRandom nucleotide sequenceWorms 501
F. hepatica, F. gigantica PCR-RFLPGel electrophoresisSSU rDNAWorms 502
F. hepatica, F. gigantica, intermediate Fasciola PCR-RFLPGel electrophoresisITS2Worms 503
F. hepatica, F. gigantica, intermediate Fasciola PCRGel electrophoresisITS2Worms 504
F. hepatica, F. gigantica LAMPVisualIGSWorms, cercariae, eggs 505
F. hepatica, F. gigantica Multiplex PCRGel electrophoresisMitochondrial DNAWorms, miracidia, eggs, stools 506
Paragonimus PCR sequencingSequenceITS2Eggs 507
P. westermani LAMPVisualITS2Sputum, pleural fluid, crabs, crayfish 380
P. westermani, Fasciolopsis buski, F. gigantica PCRGel electrophoresisITS2Worms 508
Paragonimus heterotremus, P. westermani, P. macrorchis, P. siamensis, P. harinasutai, and P. bangkokensis Real-time PCRFRET probes and MCAITS2Feces of experimentally infected cats 509
Paragonimus bangkokensis, P. harinasutai, P. heterotremus, P. macrorchis, P. siamensis, P. westermani PCR pyrosequencingSequenceITS2Metacercariae 510
  • a PCR primers are described in reference 491.

  • b PCR primers are described in reference 499.

  • c Abbreviations: RFLP, restriction fragment length polymorphism; MCA, melt curve analysis; HRM, high-resolution melt curve analysis; SSU, small subunit; ITS, internal transcribed spacer; HSP, heat shock protein; IGS, intergenic spacer region; cox1, mitochondrial cytochrome c oxidase subunit I gene; nad1, NADH dehydrogenase subunit 1; nad2, NADH dehydrogenase subunit 2; MLPA, multiple ligation-dependent probe amplification; HAT-RAPD, high-annealing-temperature random amplified polymorphic DNA; LAMP, loop-mediated isothermal amplification; FRET, fluorescence resonance energy transfer.