The rapid detection method by polymerase chain reaction for minute intestinal trematodes: Haplorchis taichui in intermediate snail hosts based on 18s ribosomal DNA

摘要

The minute intestinal trematode, Haplorchis taichui, is an important parasite species that can infect humans and other mammals. This study investigated the outbreak of H. taichui in thiarid snails in the lower part of the Chao Phraya Basin, Thailand by employing morphological and molecular-based methods. In development of a specific primer of H. taichui, the PCR reaction was conducted with no cross-reaction to their hosts and other related trematode species. The highest level of sensitivity that could be amplified was 0.50 ng/μl and this was detected with only one egg in the sample. In terms of the epidemic results, the parapleurolophocercous cercaria infected only two species of thiarid snails (Melanoides tuberculata and Tarebia granifera) with an overall prevalence of 3.80% (23/605). The process of molecular identification revealed positive results indicating that eleven from twenty-three of parapleurolophocercous cercariae specimens in the lower part of the Chao Phraya Basin were H. taichui. In conclusion, this study has developed a rapid detection method, which can discriminate H. taichui from other parapleurolophocercous cercaria in intermediate snail hosts with a high level of sensitivity. Moreover, the high proportion of H. taichui in parapleurolophocercous cercaria (47.83%) indicated that H. taichui was the dominant species of this cercarial type and could infect cyprinoid fish in the lower part of the Chao Phraya Basin leading to public health problems in this area. Thus, a specific primer could be useful in the detection and surveillance of H. taichui outbreaks in their hosts. Recognition of this has resulted in the creation of important prevention programs in these infected areas in the further study.