Development of PCR-based diagnosis of minute intestinal fluke, Haplorchis taichui

Abstract

Haplorchis taichui specific primers were designed using a high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR method and 18 arbitrary primers (Operon Technologies) to generate polymorphic DNA profiles for 13 different parasites. The H. taichui specific fragment was screened. A 256 bp HAT-RAPD marker generated from OPP-11 primer specific for H. taichui was cloned and sequenced. From the sequence data, specific primers were designed that generated a 256 bp amplicon. The minimum DNA template needed for PCR detection was 10 fg. The successful development of the H. taichui specific DNA-based detection will be beneficial in management and epidemiological control programs.