Discrimination of Neospora caninum from Toxoplasma gondii and other apicomplexan parasites by hybridization and PCR.

Kaufmann, H; Yamage, M; Roditi, I; Dobbelaere, D; Dubey, J P; Holmdahl, O J; Trees, A; Gottstein, Bruno (1996). Discrimination of Neospora caninum from Toxoplasma gondii and other apicomplexan parasites by hybridization and PCR. Molecular and cellular probes, 10(4), pp. 289-297. Elsevier

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Neospora caninum is a protozoan parasite which causes neurological problems in dogs and abortion in cattle. As N. caninum is difficult to distinguish morphologically from Toxoplasma gondii, we developed a molecular tool capable of discriminating between the two parasites. Genomic DNA was isolated from in vitro cultured N. caninum tachyzoites and cloned into a plasmid vector. Resulting colonies were subsequently screened by differential hybridization using N. caninum and T. gondii DNA. Two clones were characterized in detail: one clone, termed pNc5, was found to be specific for N. caninum whereas the second clone, pNc1, hybridized with DNA from both parasites. The sequence of pNc5 was determined and different oligonucleotide primers were designed for use in the polymerase chain reaction (PCR). A 944 bp fragment was specifically amplified from N. caninum DNA, but not from DNA extracted from T. gondii or different Sarcocystis species. Positive signals in PCR were obtained with as little as 100 pg parasite template DNA. In addition, dual PCR with primer pairs specific for N. caninum and T. gondii allowed the detection of either parasite in mixed samples.

Item Type:	Journal Article (Original Article)
Division/Institute:	05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology
UniBE Contributor:	Gottstein, Bruno
Subjects:	600 Technology > 610 Medicine & health 600 Technology > 630 Agriculture
ISSN:	0890-8508
Publisher:	Elsevier
Language:	English
Submitter:	Bruno Gottstein
Date Deposited:	23 Jul 2018 12:33
Last Modified:	05 Dec 2022 15:16
PubMed ID:	8865177
URI:	https://boris.unibe.ch/id/eprint/118711