Blocking ELISA Using Recombinant NcSRS2 Protein for Diagnosing Bovine Neosporosis

Sinnott, Francine Alves; Monte, Leonardo Garcia; Collares, Thaís Farias; Matos, Bruno Moisés de; Pacheco, Diene B.; Borsuk, Sibele; Andreotti, Renato; Hartleben, Cláudia Pinho

doi:10.1007/s00284-014-0737-y

Cited by 7 publications

(5 citation statements)

References 23 publications

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“…On the other hand, the cutoff value differentiated 74 to 80 false-negative results (92.9% specificity) (Table 3). By comparing the sensitivity and specificity parameters to the average diagnostic accuracy, we found that the chimeric protein afforded equivalent or better values than those presented in the literature (5,7,(14)(15)(16)(17)(18)(19). Moreover, the chimeric protein can discriminate infected samples from abortive diseases such as brucellosis, leptospirosis, and trypanosomiasis, which were not tested in previous articles, as well as from phylogenetically related pathogens, including T. gondii, Sarcocystis spp., and Besnoitia besnoiti.…”

Section: Discussionmentioning

confidence: 80%

“…ELISA is a high-throughput, fast, and cost-effective method that can be used to investigate a multitude of samples in fewer reactions and has been well characterized in serological diagnosis (45). The indirect fluorescent antibody test is a reference technique used to diagnosis neosporosis, although it is expensive, laborious, and less accurate than ELISA (14). An indirect ELISA with chimeric protein as the antigen was performed to measure diagnostic potential in serum samples (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Among the common serological techniques, enzyme-linked immunosorbent assay (ELISA) can be easily automated to evaluate multiple samples at low cost. Some recombinant antigens for serodiagnosis bovine neosporosis have already been described, including rNcGRA7 and rNcSAG1 (5), NcGRA7 (7), NcSRS2 (14), NcMIC10 (15), NcSAG1 (16), NcSAG4 (17), surface antigen p40 (18), and subtilisin-like serine protease 1 (19). Overall, these antigens exhibit a high potential to recognize Neospora caninum antibodies in infected animals, thereby differentiating healthy and toxoplasmosisinfected samples.…”

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confidence: 99%

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Chimeric Protein Designed by Genome-Scale Immunoinformatics Enhances Serodiagnosis of Bovine Neosporosis

et al. 2020

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Neosporosis has become a concern since it is associated with abortion in cattle. Currently, in situ diagnosis is determined through anamnesis, evaluation of the history, and perception of the clinical signs of the herd. There is no practical and noninvasive test adapted to a large number of samples, which represents a gap for the use of new approaches that provide information about infections and the risks of herds. Here, we performed a search in the Neospora caninum genome by linear B-cell epitopes using immunoinformatic tools aiming to develop a chimeric protein with high potential to bind specifically to antibodies from infected cattle samples. An enzyme-linked immunosorbent assay with the new chimeric antigen was developed and tested with sera from natural field N. caninum-infected bovines. The cross-reactivity of the new antigen was also evaluated using sera from bovines infected by other abortive pathogens, including Trypanosoma vivax, Leptospira sp., Mycobacterium bovis, and Brucella abortus, and enzootic bovine leucosis caused by bovine leukemia virus, as well as with samples of animals infected with Toxoplasma gondii. The assay using the chimeric protein showed 96.6% ± 3.4% of sensitivity in comparison to healthy animal sera. Meanwhile, in relation to false-positive results provided by cross-reactivity with others pathogens, the specificity value was 97.0% ± 2.9%. In conclusion, immunoinformatic tools provide an efficient platform to build an accurate protein to diagnose bovine neosporosis based on serum samples.

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Section: Discussionmentioning

confidence: 80%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Chimeric Protein Designed by Genome-Scale Immunoinformatics Enhances Serodiagnosis of Bovine Neosporosis

et al. 2020

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“…Nevertheless, whole tachyzoite lysate antigens (NLA) are still widely used and are considered as a standard antigen in evaluating new diagnostic candidates. In cases of cattle, specific antibodies to NcSAG1, NcSRS2, NcGRA6, and NcGRA7 [209][210][211][212][213] were detected. Furthermore, specific antibodies to NLA, especially IgG1 and IgG2, were detected in sera of infected cattle [214][215][216].…”

Section: Humoral Immune Responsesmentioning

confidence: 99%

From Signaling Pathways to Distinct Immune Responses: Key Factors for Establishing or Combating Neospora caninum Infection in Different Susceptible Hosts

Fereig

Nishikawa

2020

Pathogens

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Neospora caninum is an intracellular protozoan parasite affecting numerous animal species. It induces significant economic losses because of abortion and neonatal abnormalities in cattle. In case of infection, the parasite secretes numerous arsenals to establish a successful infection in the host cell. In the same context but for a different purpose, the host resorts to different strategies to eliminate the invading parasite. During this battle, numerous key factors from both parasite and host sides are produced and interact for the maintaining and vanishing of the infection, respectively. Although several reviews have highlighted the role of different compartments of the immune system against N. caninum infection, each one of them has mostly targeted specific points related to the immune component and animal host. Thus, in the current review, we will focus on effector molecules derived from the host cell or the parasite using a comprehensive survey method from previous reports. According to our knowledge, this is the first review that highlights and discusses immune response at the host cell–parasite molecular interface against N. caninum infection in different susceptible hosts.

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“…Dogs can be infected through three ways; ingestion of the infected tissues from the intermediate hosts, vertical transmission or consumption of the sporulated oocysts from the environment [14][15][16]. Canines, both domestic and wild are known as nal hosts of the protozoan, whereas a wide variety of mammals especially cattle and sheep, act as intermediate hosts [17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Development and application of an indirect ELISA for the Neospora caninum based on a fusion protein rSRS2- SAG1-GRA7

Yang

Opeyemi

et al. 2022

Preprint

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Introduction: Neospora caninum is an important apicomplexan parasite causing neosporosis in cattle. The disease is recognized as one of the most important cause of reproductive problems and abortion in cattle worldwide. In this context, we developed an indirect ELISA with fusion protein rSRS2-SAG1-GRA7 for the diagnosis of neosporosis in aborted dairy cattle. Results: The reaction conditions of the developed indirect ELISA were optimized. This indirect ELISA was compared to indirect immunoinfluscent assay (IFA), a total of 56 sera samples from cattle were used to evaluate the ELISA, the antibodies against N. caninum were tested by this ELISA to evaluate the prevalence of N. caninum in Ningxia. The sensitivity and specificity results of the rSRS2-SAG1-GRA7 indirect ELISA were 83.33% and 100%, the overall coincidence rate was 94.64%. N. caninum was prevalent in all the 4 cities investigated in Ningxia. The positive rate was from 16.67 to 100 %, and the mean rate was 41.64%. Conclusions: This established rSRS2-SAG1-GRA7 indirect ELISA is capable of detecting serum antibodies against N. caninum, and due to its high sensitivity and specificity, it could be applied for serological evaluation and indirect diagnosis of N. caninum infection. And we assessed N. caninum seroprevalence in aborted dairy cattle from Ningxia province of China with this rSRS2-SAG1-GRA7 indirect ELISA.

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Blocking ELISA Using Recombinant NcSRS2 Protein for Diagnosing Bovine Neosporosis

Cited by 7 publications

References 23 publications

Chimeric Protein Designed by Genome-Scale Immunoinformatics Enhances Serodiagnosis of Bovine Neosporosis

Chimeric Protein Designed by Genome-Scale Immunoinformatics Enhances Serodiagnosis of Bovine Neosporosis

From Signaling Pathways to Distinct Immune Responses: Key Factors for Establishing or Combating Neospora caninum Infection in Different Susceptible Hosts

Development and application of an indirect ELISA for the Neospora caninum based on a fusion protein rSRS2- SAG1-GRA7

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