Echinostoma revolutum: Development of a high performance DNA-specific primer to demonstrate the epidemiological situations of their intermediate hosts.

Echinostomiasis caused by the Echinostoma group, in particular E. revolutum are a significant problem for both humans and other animals. This group has a large number of morphological similarities that are difficult and time-consuming to identify. The present study aimed to develop high-performance tools for the detection of the prevalence of E. revolutum and to reveal the prevalence of E. revolutum infections in intermediate snail hosts in Lopburi province, Thailand. The snail specimens were collected by stratified sampling method and examined to collect trematodes in the larval stage. The specific primer was manually designed and based on 18 s rDNA and verified the specificity and sensitivity for use as an identification tool to compare with classical method, constructed by epidemic mapping. The overall prevalence value of E. revolutum was found to be 16.26%. Tha Luang district had the highest prevalence (70.14%), followed by Chai Badan, Phatthana Nikhom, Tha Wung, Ban Mi, Khok Samrong, Nong Muang and Sa Bot at 42%, 25.14%, 2.52%, 1.73%, 2%, 1.33% and 0.40%, respectively. With regard to the specific primer, it can amplify both cercarial and metacercarial DNA (90 pg/μl.) and discriminated E. revolutum from its hosts, other trematodes and other echinostome larvae with no cross-reactions. Therefore, the developed specific primer can be used as a species-specific identification tool with a high degree of sensitivity and specificity. Consequently, this data is important for monitoring the outbreak of E. revolutum. It can be applied for initiating surveillance programs of snail-borne diseases in both medical and veterinary studies.