Detection of Neospora caninum DNA by the polymerase chain reaction

Payne, Suzanne; Ellis, John

doi:10.1016/0020-7519(96)00030-6

Cited by 87 publications

(46 citation statements)

References 17 publications

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“…DNA from whole blood, seminal fluid and the nonsperm cell fraction were analysed separately using nested-PCR on the internal transcribed spacer (ITS1) region of N. caninum [8]. Nested-PCR was carried out using four oligonucleotides, as previously described by Buxton et al [9].…”

Section: Pcr Proceduresmentioning

confidence: 99%

Experimental neosporosis in bulls: Parasite detection in semen and blood and specific antibody and interferon-gamma responses

et al. 2007

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Aim: To investigate the presence of Neospora caninum in semen and blood, and the development of specific antibody and interferon-gamma (IFN-g) responses in experimentally infected bulls. Methods: Eight bulls were intravenously infected with 10 8 live N. caninum tachyzoites of NC-1 isolate. The presence of N. caninum in semen and blood was assessed using a nested-PCR procedure. PCR-positive semen samples were bioassayed using a BALB/c nu/nu mouse model. Specific anti-N. caninum antibody and IFN-g responses were also examined. In parallel, eight seronegative bulls were studied as non-infected controls. All bulls were monitored for 26 weeks. Results: All eight experimentally infected bulls showed N. caninum DNA in their semen and/or blood samples at some time during the course of the study. Parasite load in semen ranged from 0.1 to 14.5 parasites/ml (mean 6.0). N. caninum could not be detected in BALB/c nu/nu mice inoculated with PCR-positive semen samples. A significant increase in mean serum specific IgM antibody response to N. caninum was detected between 10 and 28 days post-infection (p.i.). Serum specific IgG, IgG1, and IgG2 antibody levels in experimentally infected bulls were significantly different after 21, 10, and 14 days p.i. as compared to controls, respectively. Specific anti-N. caninum IgG were detected in seminal plasma from infected bulls and values obtained were different from controls after 25 days p.i. Mean specific IFN-g responses in experimentally infected bulls were significantly higher than controls 3 days p.i. Conclusions: This is the first study to report the presence of N. caninum DNA in the semen and blood of experimentally infected bulls. Our observations indicate an intermittent presence of N. caninum in low numbers in semen and associated with chronic stage of the infection. This study is also the first to report the detection of anti-N. caninum IgG in seminal plasma of experimentally infected bulls. #

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Section: Pcr Proceduresmentioning

confidence: 99%

Experimental neosporosis in bulls: Parasite detection in semen and blood and specific antibody and interferon-gamma responses

et al. 2007

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“…Two PCR systems were used, i.e., classic PCR targeting the ITS1 DNA region (Payne and Ellis, 1996) and a previously described real-time PCR targeting the NC5 DNA region (Ghalmi et al, 2008). For ITS1 PCR (Payne and Ellis, 1996), an internal PCR control has been developed by amplifying invA gene of Salmonella typhimurium with the use of CI-ITS1F (59-gctgataatgaaagtgtgccggaagtattgtt-39) and CI-TS1R (59-aaataacggtgtgggaaaa cctcttcatgcgttacccag-39) primers. The resulting amplicon (240 base pairs [bp]) has been reamplified by N. caninum-specific primers NS1 (59-gctgataatgaaagtgtg-39) and SR1 (59-aaataacggtgtgggaaaa-39).…”

Section: Aborted Fetusesmentioning

confidence: 99%

“…More recently, classic (Mü ller et al, 1996;Payne and Ellis, 1996;Yamage et al, 1996), nested (Baszler et al, 1999;Paula et al, 2004), or real-time polymerase chain reaction (PCR) (Mü ller et al, 2002;Ghalmi et al, 2008) methods have been developed to detect N. caninum in dogs and cattle. Case control studies are efficient epidemiological tools used to investigate the relationship between a disease and a particular factor.…”

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confidence: 99%

Neospora caninum Is Associated With Abortion In Algerian Cattle

Ghalmi

China

Kaidi

et al. 2011

Journal of Parasitology

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Neospora caninum is a major cause of abortion in cattle worldwide. However, little information is available for Algeria. Accordingly, 799 cattle from 87 farms in the north and northeast of Algeria were enrolled in a seroepidemiological survey. An indirect fluorescence antibody test (IFAT) revealed a seroprevalence of 19.6%. The animals were divided into 3 groups according to their breed: imported European cattle, local breeds, and crossed animals (European 3 local). Seroprevalences were 16.0%, 34.3%, and 18.6% in groups 1, 2, and 3, respectively. A case control study was performed to investigate the link between global seropositivity to N. caninum and abortion risk in those cattle farms. There was a significant (P , 0.01) association between the seroprevalence against N. caninum and the occurrence of abortion in those farms (odds ratio [OR] 5 12.03). This was also observed at the individual level (OR 5 2.79). The analysis of results according to the breed revealed a significant association between seroprevalence and abortion in groups 1 and 3, but not for group 2, despite the fact that the highest seroprevalence was observed in group 2. Cerebral tissues from 5 aborted fetuses were available for histology and polymerase chain reaction (PCR). One sample was found positive both by histology and by PCR, 2 samples were positive by PCR only, and 2 samples were negative in both tests.

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“…A number of approaches have proved to be both specific and highly sensitive for analyses either of parasites grown in vitro or present in tissue samples and clinical materials, for example Toxoplasma (Burg et al, 1989;Guay et al 1993) and Neospora (Holmdahl & Mattsson, 1996;Müller et al, 1996). An attractive genomic DNA target for PCR analyses is the internal transcribed spacer 1 (ITS1) from within the ribosomal DNA (rDNA) genes (Cai et al, 1992;Holmdahl & Mattsson, 1996;Jeffries et al, 1996;Payne & Ellis, 1996). This spacer separates the 3' end of the 16S-like ribosomal RNA gene from the 5' end of the 5.8S rRNA gene within individual rDNA transcription units.…”

Section: Introductionmentioning

confidence: 99%

Development of a diagnostic PCR assay for the detection and discrimination of four pathogenic.Eimeriaspecies of the chicken

et al. 1998

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We describe a polymerase chain reaction (PCR)-based assay for the detection, identification and differentiation of pathogenic species of Eimeria in poultry. The internal transcribed spacer 1 (ITS1) regions of ribosomal DNA (rDNA) from Eimeria acervulina, E. brunetti, E. necatrix and E. tenella were sequenced and regions of unique sequences identified. Four pairs of oligonucleotide primers, each designed to amplify the ITS1 region of a single Eimeria species, were synthesised for use in the PCR assay. In tests on purified genomic DNA from all seven species of Eimeria that infect the chicken, each of the four primer pairs amplified the ITS1 region from only their respective target species. The robustness of the approach was further demonstrated by the amplification of specific DNA fragments from tissues of experimentally infected animals and from oocysts recovered from field samples. We conclude that the ITS1 regions of Eimeria species contain sufficient inter-specific sequence variation to enable the selection of primers that can be applied in PCR analyses to detect and differentiate between species. In future work they may provide excellent markers for epidemiological studies.

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Detection of Neospora caninum DNA by the polymerase chain reaction

Cited by 87 publications

References 17 publications

Experimental neosporosis in bulls: Parasite detection in semen and blood and specific antibody and interferon-gamma responses

Experimental neosporosis in bulls: Parasite detection in semen and blood and specific antibody and interferon-gamma responses

Neospora caninum Is Associated With Abortion In Algerian Cattle

Development of a diagnostic PCR assay for the detection and discrimination of four pathogenic.Eimeriaspecies of the chicken

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