A time-resolved fluorescence immunoassay for the detection of anti-Neospora caninum antibodies in sheep

Huertas-López, Ana; Martínez‐Carrasco, Carlos; Cerón, José J.; Sánchez-Sánchez, Roberto; Vázquez-Calvo, Ángela; Álvarez‐García, Gema; Martı́nez-Subiela, Silvia

doi:10.1016/j.vetpar.2019.108994

Cited by 7 publications

(4 citation statements)

References 37 publications

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“…While in dogs, specific antibody to NcSAG1, NcGRA2, NcGRA6, NcGRA7, and NcPF was detected [210,[217][218][219]. In sheep also, specific antibodies against N. caninum infection was detected as evidenced by high levels of NcGRA7 in experimentally or naturally infected animals [220]. Specific antibodies against NcSAG1, NcGRA6, and NcGRA7 in BALB/c mice [213], and NcGPI in Swiss OF1 mice [89] were detected.…”

Section: Humoral Immune Responsesmentioning

confidence: 90%

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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Section: Humoral Immune Responsesmentioning

confidence: 90%

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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“…The sample of choice is the serum, although milk has been also suggested for serological surveys of the prevalence of exposure of sheep to N. caninum [ 57 ]. At present, ELISAs have mostly replaced IFATs, and several tests have been developed or adapted specifically for ovine samples, but their reliability needs to be improved by further validation studies [ 58 ]. Although IFATs and ELISAs are the most commonly used serological tests for the diagnosis of ovine neosporosis, only a moderate agreement between them has been reported [ 59 , 60 ].…”

Section: Diagnosismentioning

confidence: 99%

“…Nevertheless, the validation of diagnostic assays is a process involving constant development and readjustment of performance characteristics for each target population [ 65 ]. This may be due to sera from experimental infections commonly being used during validation of serological tests [ 58 ], even though field sera are required due to naturally infected animals having a significantly lower immune response [ 66 ]. For this reason, the major challenge in definitively validating any serological test is collecting the sera required from the large number of naturally infected seropositive sheep from different countries.…”

Section: Diagnosismentioning

confidence: 99%

Ovine Neosporosis: The Current Global Situation

Benavides

González-Warleta

Arteche-Villasol

et al. 2022

Animals

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In the past 20 years, Neospora caninum infection in sheep has been reported in at least 31 countries worldwide from all sheep-rearing continents (Europe, Asia, the Americas, Africa, and Oceania), and its role as an abortifacient agent is becoming more evident. Most studies of ovine neosporosis have focused on its epidemiology, based primarily on serological analysis, with only a few studies investigating the actual presence of the parasite by PCR and/or IHC. Individual seroprevalence rates were highly variable between countries, and even between regions within the same country, ranging from 0.0% to 67.4% positive. Furthermore, most of the studies were not directly comparable due to differences in experimental designs, sample sizes, husbandry systems, ecological factors, and serological tests (e.g., IFAT, ELISA, MAT, Western blot). The latter, along with the scarcity of studies on the relevance of N. caninum as an abortifacient agent, may bias the perception of the importance of this disease. This review summarizes the situation of N. caninum infection in sheep using all available published studies describing natural ovine neosporosis. The epidemiology shows that ovine neosporosis is found worldwide, and it poses a relevant risk to the sustainability of sheep flocks.

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“…Fast, convenient, and fully automated detection methods are more suitable for screening animal diseases. Immunoassays can achieve A time-resolved fluorescence immunoassay (TRFIA) is a novel detection technique with high sensitivity and convenient and full automation that is highly suitable for detecting animal diseases [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling

Li,

Chen,

Liang

et al. 2024

Virol J

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Objective Canine enteric coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the main pathogens responsible for acute gastroenteritis in dogs, and both single and mixed infections are common. This study aimed to establish a double-labeling time-resolved fluorescence immunoassay (TRFIA) to test and distinguish CCV and CPV-2 diseases. Methods A sandwich double-labeling TRFIA method was established and optimized using europium(III) (Eu3+)/samarium(III) (Sm3+) chelates. CCV/CPV-2 antigens were first captured by the immobilized antibodies. Then, combined with Eu3+/Sm3+-labeled paired antibodies, the Eu3+/Sm3+ fluorescence values were detected after dissociation to calculate the CCV/CPV-2 ratios. The performance, clinical performance and methodology used for laboratory (sensitivity, specificity, accuracy and stability) testing were evaluated. Results A double-label TRFIA for CCV and CPV-2 detection was optimized and established. The sensitivity of this TRFIA kit was 0.51 ng/mL for CCV and 0.80 ng/mL for CPV-2, with high specificity for CCV and CPV-2. All the accuracy data were less than 10%, and the recovery ranged from 101.21 to 110.28%. The kits can be temporarily stored for 20 days at 4 °C and can be stored for 12 months at temperatures less than − 20 °C. Based on a methodology comparison of 137 clinically suspected patients, there was no statistically significant difference between the TRFIA kit and the PCR method. Additionally, for CCV detection, the clinical sensitivity was 95.74%, and the clinical specificity was 93.33%. For CPV-2 detection, the clinical sensitivity was 92.86%, and the clinical specificity was 96.97%. Conclusion In this study, a double-label TRFIA kit was prepared for CCV and CPV-2 detection with high laboratory sensitivity, specificity, accuracy, stability, clinical sensitivity and specificity. This kit provides a new option for screening/distinguishing between CCV and CPV-2 and may help improve strategies to prevent and control animal infectious diseases in the future.

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A time-resolved fluorescence immunoassay for the detection of anti-Neospora caninum antibodies in sheep

Cited by 7 publications

References 37 publications

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

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

Ovine Neosporosis: The Current Global Situation

Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling

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