Standardization of an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies anti-Corynebacterium pseudotuberculosis in sheep

Nassar, Alessandra Figueiredo de Castro; Miyashiro, Simone; Gregori, Fábio; Piatti, Rosa Maria; Daniel, Gabriela Terezinha; Gregory, Lílian

doi:10.1016/j.smallrumres.2013.10.016

Cited by 7 publications

(5 citation statements)

References 11 publications

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“…The advantages are cost-benefit, ease of application, and acceptable sensitivity and specificity (OREIBY, 2015). Nassar et al (2014) reported other advantages like shorter test time, ease of antigen preparation, and the use of small quantity of inputs, make the use of this test possible for disease diagnosis in the epidemiological studies and for commercial purposes. Thus, in addition to facilitating CL diagnosis in herds, ELISA contributes to disease control in situations where clinical symptoms are not evident, in turn becoming an important emerging tool to be used in the implementation of disease-control programs in Brazil.…”

Section: Discussionmentioning

confidence: 99%

Seroprevalence of caseous lymphadenitis in goats sold in an animal fair in the northeastern semi-arid region of Brazil

Alves

Farias

Lima³

et al. 2018

SCA

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Goat farming in the northeastern regions of Brazil plays an important role in the socio-economic functions of the country. However, high rates of morbidity and mortality occur in the animals, caused primarily by infectious diseases like caseous lymphadenitis, which is widespread in goat herds; this causes serious economic losses to the farming business. Although events such as animal fairs are common in the region, wherein most of the goats and sheep are sold, the risk of disease transmission remains, since the health condition of the animal is unknown. The aim of this study was to determine the frequency of anti-Corynebacterium pseudotuberculosis antibodies present in the goats sold at the animal fair of Tabira -PE, Northeastern semi-arid. Serum samples from 233 goats were collected from the period of November 2014 to June 2015. The diagnosis of infection by C. pseudotuberculosis was made using the indirect ELISA technique. The number of goats seropositive for C. pseudotuberculosis was identified as 87 (37.34%; 95% CI = 31.38 -43.71%). Of the nine flocks evaluated, five (55.55%) were positive. Thus, it is suggested that infection by C. pseudotuberculosis is widespread in the goats sold at the animal fairs in the semi-arid Northeastern, reinforcing the need for disease diagnosis in goat herds in the region for the possible implementation of disease control programs. Key words: Agglomerations. Caseous lymphadenitis. Goats. Serology. ResumoA caprinocultura se destaca na região Nordeste do Brasil por desempenhar importante função socioeconômica. Contudo altas taxas de morbidade e mortalidade ocorrem nas propriedades causadas principalmente por doenças infecciosas, destacando-se a linfadenite caseosa, que se encontra amplamente difundida nos rebanhos caprinos, acarretando sérios prejuízos econômicos à atividade. Eventos do tipo

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Section: Discussionmentioning

confidence: 99%

Seroprevalence of caseous lymphadenitis in goats sold in an animal fair in the northeastern semi-arid region of Brazil

Alves

Farias

Lima³

et al. 2018

SCA

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“…Original Research samples were diluted 1:200 in PBS and added to the plates in duplicate 18 . The plates were subsequently incubated for 1 h at 37 °C and washed three times with PBS-T. Then, 100 µl of the corresponding isotype (anti-IgG1, anti-IgG2a, anti-IgG2b, or anti-IgM) was added and diluted to 1:1,000 in PBS.…”

Section: Evaluation Of the Humoral And Cellular Immune Responses Indumentioning

confidence: 99%

“…The cells were then cultivated until the log phase for 24 h in BHI medium. Then, they were centrifuged, washed three times with PBS, and finally, sonicated for 12 min at 60 Hz 18 . The protein concentration was determined using the Bradford method and adjusted as required.…”

Section: Evaluation Of the Humoral And Cellular Immune Responses Indumentioning

confidence: 99%

Evaluation of the aroA mutant of Corynebacterium pseudotuberculosis in cellular and murine models

et al. 2016

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Caseous lymphadenitis of small ruminants causes economic losses worldwide. To date, no effective vaccine has been developed against the causative agent of this disease, Corynebacterium pseudotuberculosis. The objective of the present work was to evaluate an aroA mutant gene strain of C. pseudotuberculosis in cellular and murine models, for attenuation and the ability to stimulate an immune response. The intracellular survival of the aroA mutant strain and the wild type strain (WT) of C. pseudotuberculosis was evaluated in J774A.1 murine macrophages using a multiplicity of infection (MOI) of 1:1 with the following infection times: 30 min, and 1, 2, 4, 8, 12, and 24 h. The largest difference in the intracellular survival of the mutant was observed 30 min post-infection. After subcutaneous skin vaccination, the subcutaneous lesion progression observed on the 14 th day was more severe in those animals that were vaccinated with the WT strain. An analysis of the residual virulence in the murine model did not reveal any bacteria in mice vaccinated with the aroA strain on day 28 post-vaccination. Mice vaccinated with the mutant showed 50 % protection against the intraperitoneal challenge, exceeding that of the control group (41.67 %). We conclude that the virulence of the aroA mutant was significantly attenuated in both cellular and murine models according to the residual virulence detected in mice. However, vaccination with the mutant failed to confer at least 80 % protection, which is desirable for an immunogen. Hence, this study contributes to the knowledge of the immune response against Corynebacterium pseudotuberculosis.

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“…The difficulty in accurately identifying subclinical cases of CLA favours the spread of the disease within and between herds. The screening and elimination of subclinical animals is an important measure in disease control programmes (Farias et al, 2019;Nassar et al, 2014). ELISA (enzyme-linked immunosorbent assay) is an economical and simple method to perform as a routine technique in CLA diagnosis; however, ELISA tests are not being used in the control and eradication programmes of CLA, either because they present low sensitivity and specificity (Menzies, Hwang, & Prescott, 2004), have not been fully tested or have not been prepared for both species, sheep and goat (Hoelzle et al, 2013;Oreiby Atef, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…ELISA (enzyme-linked immunosorbent assay) is an economical and simple method to perform as a routine technique in CLA diagnosis; however, ELISA tests are not being used in the control and eradication programmes of CLA, either because they present low sensitivity and specificity (Menzies, Hwang, & Prescott, 2004), have not been fully tested or have not been prepared for both species, sheep and goat (Hoelzle et al, 2013;Oreiby Atef, 2014). In largescale control and eradication programmes, or for the detection of animals with unapparent infection and, consequently, contributing to preventing the dispersion of the disease, ELISA would be the most suitable technique (Hoelzle et al, 2013;Menzies et al, 2004;Nassar et al, 2014;Oreiby et al, 2015;Oreiby Atef, 2014). Very few studies have evaluated the best diagnostic technique to detect subclinical cases of CLA (Barral et al, 2019), but the use of serodiagnosis as a tool for the control and eradication of CLA in goat farms has been successfully implemented in some cases (Derckeson et al, 2000;Kaba, Kutschke, & Gerlach, 2001).…”

Section: Introductionmentioning

confidence: 99%

Utility assessment of an Enzyme‐linked immunosorbent assay for detection of subclinical cases of caseous lymphadenitis in small ruminant flocks

Costa

Huerta

Galán‐Relaño

et al. 2020

Veterinary Medicine & Sci

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The actual prevalence of CLA (caseous lymphadenitis) in small ruminant flocks is underestimated in many countries, and because it is not a notifiable disease, it will continue to spread without data and information about its real economic impact. The difficulty in the accurate identification of the causative agent in internal subclinical cases allows the disease to spread within and between flocks. This research intends to assess the utility of an ELISA (enzyme‐linked immunosorbent assay) test in the detection of internal subclinical cases of CLA in farms and to simultaneously add data on the seroprevalence of the disease in Portugal. Sera from 756 small ruminants, 70% sheep (528/756) and 30% goats (228/756) were screened for antibodies against Corynebacterium pseudotuberculosis using the ELISA technique based on a recombinant phospholipase D (ELITEST CLA # CK105A®). The animals showing internal lesions (n ꞊ 58) were sampled for the identification of the aetiological agent. In this investigation, the prevalence of CLA was 34% (258/756), with the ELISA test showing a low specificity (78%) and high sensitivity (100%). The proof was able to detect 57% (13/23) of subclinical cases of CLA confirmed by postmortem examination and conventional PCR (polymerase chain reaction). The results also reveal that goats have a higher propensity for the disease, and dairy farms and non‐extensive production units appear to be more susceptible to CLA. This research clarifies an actual problem and pointed out the importance of CLA in small ruminant herds in Portugal. Finally seems to demonstrate that the ELISA test is a good diagnostic tool for use in CLA eradication programmes.

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Standardization of an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies anti-Corynebacterium pseudotuberculosis in sheep

Cited by 7 publications

References 11 publications

Seroprevalence of caseous lymphadenitis in goats sold in an animal fair in the northeastern semi-arid region of Brazil

Seroprevalence of caseous lymphadenitis in goats sold in an animal fair in the northeastern semi-arid region of Brazil

Evaluation of the aroA mutant of Corynebacterium pseudotuberculosis in cellular and murine models

Utility assessment of an Enzyme‐linked immunosorbent assay for detection of subclinical cases of caseous lymphadenitis in small ruminant flocks

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