Specificity assay for assessment of duplex PCR using DNA template from different species, visualized on 1% agarose stained with ethidium bromide. M, 1-kb ladder marker; (−), negative control (no DNA); Fh (+) and Fg (+), F. hepatica and F. gigantica positive controls (100 ng DNA template in each case). Lanes: 1, C. sinensis (CsND); 2, O. viverrini (OvBD); 3, F. gigantica (FspT4); 4, F. hepatica (FhBe); 5, H. pumilio (HpMcND); 6, F. gigantica (FspCB1); 7, H. taichui (HTA1); 8, F. gigantica (FgigTL, adult); 9, Stellantchasmus falcatus (SfQN1); 10, Echinochasmus japonicus (EcPT); 11, Fasciolopsis buski (FbL2); 12, Taenia solium (TsoVN); 13, F. gigantica (FgigTL, eggs).