This study aimed to investigate the prevalence of cercarial infections in freshwater snails collected from the Chai Nat province of Central Thailand. Moreover, we aimed to develop a duplex PCR technique, using the ITS2 and candidate MT-ND1 gene, to determine the dissemination of H. taichui and H. pumilio, respectively. Six types of cercariae were discovered with an overall prevalence of 4.71% (59/1,252). The parapleurolophocercous cercariae were demonstrated to be the dominant type, infecting only Melanoides tuberculata snails. The duplex PCR was optimized for specific amplification of ITS2 for H. taichui (115 bp) and MT-ND1 for H. pumilio (335 bp). Both specific primers confirmed the specificity of the duplex PCR reaction, with no cross-reactivity with other heterophyids or related species. In addition, this duplex PCR could be used to detect co-infection at a concentration of 3.0 ng/µL. For the molecular identification, 9 of 22 parapleurolophocercous cercaria specimens in Chai Nat province generated the specific DNA fragment of H. pumilio. These results proved that the MT-ND1 gene is a species-specific method for heterophyid detection and provides a rapid method for identification based on larval and adult stages of H. taichui and H. pumilio in their intermediate and/or definitive hosts in the infected area.

这项研究旨在调查从泰国中部柴纳省采的淡水蜗牛中宫颈感染的患病率。此外，我们旨在开发一种双重PCR技术，使用ITS2和候选MT-ND1基因，分别确定taichui和pumilio的传播。发现了六种尾c，总患病率为4.71％（59 / 1,252）。已证实副猪尾肺尾be是主要类型，仅感染结核分枝杆菌。双工PCR被用于ITS2的特异性扩增优化H. taichui（115碱基对）和MT-ND1为H. pumilio（335 bp）。两种特异性引物均证实了双链PCR反应的特异性，且与其他杂种或相关物种没有交叉反应。此外，该双链PCR可用于检测3.0 ng / µL浓度的共感染。为了进行分子鉴定，柴纳省的22例胸膜肺尾部尾c样本中有9例产生了H. pumilio的特定DNA片段。这些结果证明，MT-ND1基因是heterophyid检测物种特异性方法，并提供基于的幼虫和成虫阶段用于识别的快速方法H. taichui和H. pumilio在它们的中间和/或最终宿主中感染区。