The Development and Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Method for the Detection of Spirometra mansoni in Dogs

<i>Spirometra mansoni</i> is a zoonotic parasite that inhabits the intestines of dogs and cats. The plerocercoids (spargana) parasitize several vertebrates, including humans, resulting in a food-borne zoonosis known as sparganosis. In this study, it has been established that a LAMP assay can detect <i>S. mansoni</i> eggs in dog feces. A total of 97 fecal samples were collected from Changsha City, Hunan Province. The fecal DNA was extracted before designing primers for LAMP based on the <i>S. mansoni</i> cox1 gene. The specificity of this method was verified by PCR using LAMP outer primers or inner primers and nested PCR with <i>S. mansoni</i>-specific cox1 primers. DNA samples from five control dog worms were analyzed using the LAMP assay to evaluate the specificity. The detection rate of LAMP for <i>S. mansoni</i> eggs was 70.21% in stray dogs. PCR and nested PCR produced specific bands on agarose gel electrophoresis consistent with the expected length. When the LAMP assay was conducted using <i>S. mansoni</i>-infected samples, negative samples, and genomic DNA from control worms, only the <i>S. mansoni</i>-infected samples showed a typical ladder pattern. The samples were stained with SYBR Green I, and only the <i>S. mansoni</i>-infected samples had a fluorescent signal. In addition, compared with PCR and microscope, LAMP method can detect eggs in the shortest infection days, and its detection rate was higher than that of PCR. These results suggest that the established LAMP method have many advantages in detecting <i>Spirometra mansoni</i>.