Development of real-time polymerase chain reaction assays allowing molecular detection ofEchinococcus felidis, Echinococcus granulosussensu stricto, andEchinococcus canadensisin carnivore feces samples, animal and human hydatid cyst material from Uganda and Kenya

Abstract

___________________________________________________________________________First evaluations on field samples, including carnivore feces, animal and human hydatid cyst material from Uganda and Kenya, showed specific amplification of two target regions of the mitochondrial genome of Echinococcus species according to melt and high-resolution melt curve analyses of the developed real-time PCR assays. Consecutive sequencing of PCR products revealed that, apart from Echinococcus felidis, sequences of two other tapeworm species, Echinococcus granulosus sensu stricto and Echinococcus canadensis, which are also endemic in East Africa, were detected by the developed real-time PCR assays. ___________________________________________________________________________ IntroductionThe aim of the present study was to develop a rapid, sensitive, and safe method for molecular detection of Echinococcus felidis, and possibly, related tapeworm species. We first investigated nucleic acid extraction methods suitable for isolating templates directly from fecal samples of carnivores and hydatid cyst material of intermediate hosts in the field without tedious laboratory based enrichment methods, such as digestion or flotation methods. Then, we developed real-time PCR assays which target two different mitochondrial genome regions of E. felidis.

Development of Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick Assay for a Rapid and Sensitive Detection of Cystic Echinococcosis in Livestock in Kenya

“…Based on these features, the LAMP technology has gained momentum for nucleic acid amplification and has been widely used in various fields including biomedical, agriculture, veterinary, aquaculture, food, and environmental sciences [18]. e LAMP technique has been evaluated as a molecular diagnostic tool for a variety of pathogenic microorganisms including E. granulosus [19][20][21].…”