Loop-Mediated Isothermal Amplification Method for Differentiation and Rapid Detection of Taenia Species

Rapid detection and differentiation ofTaenia species are required for the control and prevention of taeniasis and cysticercosis in areas where these diseases are endemic. Because of the lower sensitivity and specificity of the conventional diagnosis based on microscopical examination, molecular tools are more reliable for differential diagnosis of these diseases. In this study, we developed and evaluated a loop-mediated isothermal amplification (LAMP) assay for differential diagnosis of infections withTaenia species with cathepsin L-like cysteine peptidase (clp) and cytochromec oxidase subunit 1 (cox1) genes. LAMP with primer sets to the cox1 gene could differentiate between three species, and LAMP with primer sets to the clp gene could differentiateTaenia solium fromTaenia saginata /Taenia asiatica . Restriction enzyme digestion of the LAMP products from primer set Tsag-clp allowed the differentiation ofTaenia saginata fromTaenia asiatica . We demonstrated the high specificity of LAMP by testing known parasite DNA samples extracted from proglottids (n = 100) and cysticerci (n = 68). LAMP could detect one copy of the target gene or five eggs ofT. asiatica andT. saginata per gram of feces, showing sensitivity similar to that of PCR methods. Furthermore, LAMP could detect parasite DNA in all taeniid egg-positive fecal samples (n = 6). Due to the rapid, simple, specific, and sensitive detection ofTaenia species, the LAMP assays are valuable tools which might be easily applicable for the control and prevention of taeniasis and cysticercosis in countries where these diseases are endemic.