Development and preliminary evaluation of two novel PCR assays for improved detection of Taenia saginata and Taenia solium eggs

Taenia saginata and Taenia solium eggs are distributed in the environment through open defecation and wastewater. This research addressed the critical need for sensitive and specific molecular methods to investigate environmental contamination with T. saginata and T. solium in challenging and complex environmental settings. Two polymerase chain reaction (PCR) assays were developed, one explicitly targeting T. saginata and the other designed to detect both T. saginata and T. solium simultaneously. Primers were designed to target the mitochondrial cox1 gene. The preliminary sensitivity of the PCRs was evaluated using DNA extracted from taeniid eggs. The sensitivity of two newly developed PCRs was compared with that of three previously used PCRs, using ten batches of eggs from T. saginata proglottids of varying ages, ranging from 2 to 89 weeks. From each batch, one, two, and five eggs were collected in triplicate using a micro pick-and-place system. Two and five eggs from one batch of T. solium eggs were also tested. For specificity, PCRs were tested against genetically similar parasites, including Taenia hydatigena , Echinococcus spp., and Hymenolepis spp. The analysis yielded a sensitivity of 92% for the T. saginata -specific PCR and 87% for the Taenia PCR. Taenia hydatigena , E. multilocularis , E. granulosus , E. canadensis, H. diminuta, and H. nana DNA did not result in any amplification with either of the new methods. The study validated the potential of two new PCRs to investigate environmental contamination.