ELISAs based on rNcGRA7 and rNcSAG1 antigens as an indicator of Neospora caninum activation

Hiasa, Jun; Kohara, Junko; Nishimura, Maki; Xuan, Xuenan; Tokimitsu, Hiromi; Nishikawa, Yoshifumi

doi:10.1016/j.vetpar.2012.01.036

Cited by 22 publications

(27 citation statements)

References 24 publications

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“…It has been reported that titers of anti-NcGRA7 antibody in cows with a history of abortion are significantly higher than in non-aborting cows that were infected with N. caninum [9]. In addition, the frequency of anti-NcGRA7 antibody-positive individuals was higher among cows with a recent history of N. caninum -associated abortion than among cows that had experienced N. caninum -associated abortion more than 30 days previously [8]. In contrast, the frequency of anti-NcSAG1 antibody-positive individuals was similar between the two groups [8].…”

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confidence: 99%

“…In addition, the frequency of anti-NcGRA7 antibody-positive individuals was higher among cows with a recent history of N. caninum -associated abortion than among cows that had experienced N. caninum -associated abortion more than 30 days previously [8]. In contrast, the frequency of anti-NcSAG1 antibody-positive individuals was similar between the two groups [8]. These phenomena might reflect the fact that anti-NcGRA7 antibodies are observed only during acute infection, including those infections arising out of recrudescence from a latent infection [1].…”

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confidence: 99%

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Prevalence and Dynamics of Antibodies against NcSAG1 and NcGRA7 Antigens of <i>Neospora caninum</i> in Cattle during the Gestation Period

Takashima

Takasu

Yanagimoto

et al. 2013

The Journal of Veterinary Medical Science

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Bovine abortion caused by the Apicomplexan parasite Neospora caninum is a major economic problem in the livestock industry worldwide. Our study measured the prevalence and temporal changes in levels of antibodies specific for two N. caninum derived antigens, NcSAG1 and NcGRA7, to determine an appropriate strategy for serodiagnosis. Using an enzyme-linked immunosorbent assay (ELISA), blood samples showed that 71 cows out of 129 were positive for anti-NcSAG1 antibodies and that only nine cows were positive for anti-NcGRA7 antibodies. By longitudinal sampling, it was revealed that positive and negative antibody conversion occurred frequently for anti-NcGRA7, but that anti-NcSAG1 antibodies persisted for a long-term. These results indicate the usefulness of measuring anti-NcSAG1 antibody levels for the detection of chronically infected cows. Twelve cows showed positive seroconversion during pregnancy, nine of which showed seropositivity for anti-NcGRA7 antibody at the sixth and/or seventh month of pregnancy; serum samples were not obtained from the remaining three cows during this period. Therefore, the optimal time for detection of anti-NcGRA7 antibodies appears to be between the fifth and eighth month of pregnancy.

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confidence: 99%

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confidence: 99%

Prevalence and Dynamics of Antibodies against NcSAG1 and NcGRA7 Antigens of <i>Neospora caninum</i> in Cattle during the Gestation Period

Takashima

Takasu

Yanagimoto

et al. 2013

The Journal of Veterinary Medical Science

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“…Currently, several antigenic proteins, such as NcSAG1t and NcGRA7, are used in serodiagnostic assays for bovine neosporosis (21). The present study illustrates the diagnostic potential of NcSUB1, which contains five copies of tandem repeat elements, to complement or improve current serodiagnosis.…”

Section: Discussionmentioning

confidence: 69%

“…The ELISA was performed as described previously (20,21). The GST-fused NcSUB1t, NcSUB1tr, N54, and NcSAG1t recombinant proteins and the GST control were diluted in coating buffer (50 mM carbonate-bicarbonate buffer [pH 9.6]) at a final concentration of 0.1 M. Each well of a flat-bottomed 96-well microtiter plate (Nunc, Denmark) was coated with 50 l of the corresponding protein and incubated overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Identification of a Highly Antigenic Region of Subtilisin-Like Serine Protease 1 for Serodiagnosis of Neospora caninum Infection

Ybañez

Terkawi

Kameyama

et al. 2013

Clin. Vaccine Immunol.

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Neospora caninum is an apicomplexan parasite that causes abortion in cattle; hence, accurate diagnosis of this pathogen is important to the cattle farming industry. Our previous proteomics and immunoscreening analyses revealed that the N. caninum subtilisin-like serine protease 1 (NcSUB1) has potential as a serodiagnostic tool for Neospora. Consequently, we expressed two fragments containing five NcSUB1 tandem repeat copies covering amino acids (aa) 524 to 843 (NcSUB1t) and 555 to 679 (NcSUB1tr) to identify the antigenic regions. The serodiagnostic performances of NcSUB1t and NcSUB1tr were compared with that of N54, which contains a single copy of the repeats (aa 649 to 784), and with the truncated NcSAG1 (NcSAG1t), which lacks a signal peptide and C-terminal hydrophobic regions, as a positive reference. Serum samples from N. caninum experimentally infected cattle and mice and cattle from a farm with confirmed cases of Neospora abortion were tested by enzyme-linked immunosorbent assay (ELISA) with the four antigens. In the N. caninum experimentally infected cattle, the highest IgG1 antibody titers were detected against NcSUB1t, while specific IgG1 antibodies were detectable from 16 days postinfection (dpi), with levels peaking at 36 dpi for all of the antigens. On the other hand, the levels of anti-NcSUB1 IgG2 antibodies were lower than those of anti-SAG1t IgG2 antibodies. The ELISA with NcSUB1t and NcSUB1tr had good sensitivity (94.59 to 95.95%) and specificity (80 to 100%) with bovine serum field samples compared to NcSAG1t and showed no cross-reactions with sera from Toxoplasma gondii experimentally infected mice. Moreover, IgG antibodies against NcSUB1t were detected during parturition in the NcSAG1t antibody-positive cattle, and NcSUB1t-specific antibody transfer was observed from a mother to her calf. Our results show that the NcSUB1 tandem repeat is potentially useful for serodiagnosis of N. caninum.

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“…The results indicated that the production of the anti-NcGRA7 antibody was up-regulated in aborting cows, which could be of interest in the diagnoses of cows which will suffer abortion. In a more recent study, the use of recombinant NcGRA7 and NcSAG1 antigens as an indicator of Neosporaactivation was reported (Hiasa et al, 2012). These results are promising for a specific serological marker linked to abortion in cattle.…”

Section: Searching For Antibody Markers Of Abortionmentioning

confidence: 91%

Markers related to the diagnosis and to the risk of abortion in bovine neosporosis

Almerı́a

López‐Gatius

2015

Research in Veterinary Science

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ELISAs based on rNcGRA7 and rNcSAG1 antigens as an indicator of Neospora caninum activation

Cited by 22 publications

References 24 publications

Prevalence and Dynamics of Antibodies against NcSAG1 and NcGRA7 Antigens of <i>Neospora caninum</i> in Cattle during the Gestation Period

Prevalence and Dynamics of Antibodies against NcSAG1 and NcGRA7 Antigens of <i>Neospora caninum</i> in Cattle during the Gestation Period

Identification of a Highly Antigenic Region of Subtilisin-Like Serine Protease 1 for Serodiagnosis of Neospora caninum Infection

Markers related to the diagnosis and to the risk of abortion in bovine neosporosis

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