A novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNA

Background: neospora caninum is a cyst-forming,coccidian parasite which is known to cause neurological disorders in dogs and abortion and neonatal mortality in cows and other livestock. this study reports the development of a loop-mediated isothermal amplification (lamp) assay based on the neospora caninum nc-5 gene and compares its efficacy for detecting dna to that of a semi-nested pcr test. results: six primers were designed based on the nc-5 repeat region of n. caninum. specific lamp primers led to successful amplification of n. caninum dna at 63 °c in 30 min. the lamp assay was highly specific (i.e. it did not reveal cross-reactivity with other parasite species) and had a low n. caninum plasmid dna limit of detection (1 fg),which is ten times higher than that for the semi-nested pcr. lamp applicability was evaluated using a set of naturally-infected samples (59 from canine faeces and five from bovine abortions). thirty-nine percent (25/64) of the naturally-infected samples were positive for n. caninum dna by lamp and 36% (23/64) by semi-nested pcr. however,the lamp assay is much faster to perform than semi-nested pcr and provides results in 30 min. conclusion: the optimized reaction conditions described in this study resulted in a sensitive,specific and rapid technique for detecting n. caninum dna. considering the advantages of lamp for detecting n. caninum dna,further assays aimed at testing its usefulness on a wider range of field samples are recommended. © 2017 the author(s).