A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections

Abstract

Background Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction. Methods Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay. Results A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously. Conclusions This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program.

Molecular Phylodiagnosis of Taenia hydatigena from Stray Dogs in Erbil Province-Kurdistan Region/Iraq

“…These may be due to no health control measures and their habits exposed to natural infection more than owned dogs. The prevalence rate (32.54%) by molecular method (PCR), these agreed with (Zhu et al, 2019), who showed that no cross-reactivity was observed with other tapeworms inhabiting the small intestine as well as with each other. The minimum detected DNA ranged from 0.1 to 1 ng, low enough to produce results in case of low DNA yield (Mirbadie et al, 2019).…”

“…It is responsible for major medical and economic losses in animals (Ulziijargala et al, 2020), due to widespread parasite that can infect a wide range of herbivorous animals with its larval stage, Cysticercus tenuicollis (Myczkaa et al, 2019;AbouLaila et al, 2020). Furthermore, adult worms parasitize mostly present in small intestine of canids and produce eggs, which are infective to intermediate hosts (Mirbadie et al, 2019;Zhu et al, 2019). Taeniid tapeworms associated with the sheepdog life cycle.…”

“…statistical computer algorithms have radically altered how organisms are classified and how evolutionary relationships from higher taxa to species and populations can be reconstructed (Nakao et al, 2013;Zhu et al, 2019). In general, a few studies conducted mostly using partial mitochondrial cytochrome c oxidase subunit 1 (cox1) and nad1 gene sequences have reported considerable levels of genetic variation among T. hydatigena populations from different geographical regions and hosts (Rostami et al, 2013;Omar et al, 2016;Muku et al, 2020).…”

Molecular Confirmation of Massive Taenia pisiformis Cysticercosis in One Rabbit in Poland

“…Transmission of this parasite is sustained by the predator-prey relationship of the definitive and intermediate hosts. T. pisiformis infections often cause much more severe damage in intermediate hosts than in definitive hosts, where infections are mostly subclinical [ 26 ].…”

Identification and Expression Profiling of Circulating MicroRNAs in Serum of Cysticercus pisiformis-Infected Rabbits

“…The Dot-ELISA diagnostic method has been reported for detection of C. pisiformis, but has high false positive rate limit applicability [9]. Multiplex PCR has been studied as a diagnostic method for T. pisiformis in dog feces, but cannot be used for the diagnosis of rabbit cysticercosis [10]. Thus, there is an urgent need to develop a new method for early diagnosis for rabbit in order to reduce the losses due to cysticercosis.…”