Development and diagnostic validation of an ELISA based on an antigenic mixture for the detection of bovine tuberculosis

Griffa, Natanael; Moyano, Roberto Damián; Canal, A.M.; Travería, Gabriel Eduardo; Santangelo, María de la Paz; Alonso, Natalia; Romanó, Maria

doi:10.1016/j.tvjl.2020.105426

Cited by 11 publications

(13 citation statements)

References 21 publications

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“…Two different antigens for indirect enzyme-linked immunosorbent assays (ELISA) were used for the accurate detection of specific antibodies in sera of MAP-infected and healthy animals: PPA-ELISA and polyprotein-ELISA. The detection of antibodies of M. bovis in sera from infected and healthy animals was performed with a validated bTB ELISA that was previously used [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

“…Finally, infection in the 35 bTB animals was confirmed according to necropsy performed in slaughterhouses authorized by SENASA. The tissues were tested by polymerase chain reaction (PCR), by amplifying the IS 6110 , which is an insertion sequence specific to organisms in the Mycobacterium tuberculosis complex [ 37 ]. These 35 animals belong to a herd that has been PTB-free for more than 15 years.…”

Section: Methodsmentioning

confidence: 99%

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Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein

Moyano

Romero

Olivieri

et al. 2021

Veterinary Medicine International

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Bovine paratuberculosis (PTB) is caused by Mycobacterium avium subsp. paratuberculosis (MAP). The optimization of detection tests specific for MAP is crucial to improve PTB control. In this work, we aimed to develop and validate a diagnostic tool based on an ELISA to specifically detect anti-MAP antibodies from bovine serum samples. For that purpose, we designed a recombinant polyprotein containing four specific antigens from MAP and optimized the ELISA. The validation consisted of the assessment of 10 sera from PTB-infected and healthy bovines with different OD values. The diagnostic performance of the polyprotein-ELISA was evaluated by testing 130 bovine serum samples (47 healthy, 48 MAP-infected, and 35 M. bovis-infected bovines). The ELISA using the polyprotein yielded an area under the ROC curve (AUC) of 0.9912 (95% CI, 0.9758–1.007; P < 0.0001). Moreover, for this ELISA, the cut-off selected from the ROC curve based on the point with a sensitivity of 95.56% (95% CI, 0.8485–0.9946) and specificity of 97.92 (95% CI, 0.8893–0.9995) was 0.3328. Similar results were obtained with an ELISA using the commercial Paratuberculosis Protoplasmatic Antigen (PPA). However, the ELISA with the polyprotein antigen showed a better performance against sera from animals infected with Mycobacterium bovis compared to the ELISA with PPA: lower cross-reactivity (2.85% versus 25.71%). These results demonstrate a very low cross-reactivity of the polyprotein with antibodies present in serum samples from animals infected with M. bovis. The designed polyprotein and the validated ELISA could be very useful for the specific identification of MAP-infected animals in herds.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein

Moyano

Romero

Olivieri

et al. 2021

Veterinary Medicine International

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“…Also, Souza et al applied a recombinant chimera ELISA antigen of ESAT-6, MPB70, and MPB83 on sera obtained 7 days after a comparative TST, the sensitivity and specificity of the ELISA were 79.5%, and 75.5%, respectively [ 84 ]. More recently, Griffa et al used an antigenic mixture from a total extract of the reference strain AN5 and were able to confirm the M. bovis infection of 83.7% of animals that were ELISA positive 15–17 days after a negative TST, by histopathology and PCR [ 85 ]. In the same study, the specificity was 95.95% and the authors suggested the detection of antibodies of M. bovis within weeks after TST as a rapid and inexpensive way to improve bTB control.…”

Section: Diagnosis Of Btbmentioning

confidence: 99%

Review on Bovine Tuberculosis: An Emerging Disease Associated with Multidrug-Resistant Mycobacterium Species

Borham¹,

Oreiby

El-Gedawy³

et al. 2022

Pathogens

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Bovine tuberculosis is a serious infectious disease affecting a wide range of domesticated and wild animals, representing a worldwide economic and public health burden. The disease is caused by Mycobacteriumbovis and infrequently by other pathogenic mycobacteria. The problem of bovine tuberculosis is complicated when the infection is associated with multidrug and extensively drug resistant M. bovis. Many techniques are used for early diagnosis of bovine tuberculosis, either being antemortem or postmortem, each with its diagnostic merits as well as limitations. Antemortem techniques depend either on cellular or on humoral immune responses, while postmortem diagnosis depends on adequate visual inspection, palpation, and subsequent diagnostic procedures such as bacterial isolation, characteristic histopathology, and PCR to reach the final diagnosis. Recently, sequencing and bioinformatics tools have gained increasing importance for the diagnosis of bovine tuberculosis, including, but not limited to typing, detection of mutations, phylogenetic analysis, molecular epidemiology, and interactions occurring within the causative mycobacteria. Consequently, the current review includes consideration of bovine tuberculosis as a disease, conventional and recent diagnostic methods, and the emergence of MDR-Mycobacterium species.

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“…As the diagnosis of mycobacterial infections, both those caused by Mycobacterium tuberculosis complex (MTBC) and NTM, still presents difficulties, there is a need to improve diagnostic tests [ 4 , 5 ]. One such difficulty is presented by the influence of cross-reactive immune responses towards different mycobacteria [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

The Risk of False-Positive Serological Results for Paratuberculosis in Mycobacterium bovis-Infected Cattle

et al. 2021

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Both bovine tuberculosis (BTB) and paratuberculosis (paraTB) continue to cause significant economic losses in cattle breeding; in addition, their etiological agents have zoonotic potential. Although the diagnostics of both diseases are still being improved, problems still remain, such as the potential for cross-reactivity to the antigens used in tests. The aim of the present study was to confirm whether animals known to harbor Mycobacterium bovis antibodies are at increased risk of yielding positive results in paraTB serotesting and, additionally, to verify the accuracy of three commonly used methods for confirming M. bovis infection: ELISA, the tuberculin skin test (TST), and the presence of gross lesions. Material was collected from 98 dairy cattle suspected of BTB due to TST-positive results. During postmortem examination, gross lesions were assessed visually. Blood, lymph nodes, and TB-suspected organs were collected. Serum was obtained from the collected blood and tested serologically for TB and paraTB. The tissues underwent standard microbiological testing for M. tuberculosis complex. Among the 98 TST-positive individuals, tuberculous gross lesions were detected in 57 (58.1%), MTBC were isolated in 83 (84.7%), and the ELISA test was positive for 21 (21.4%). None of the lesions characteristic for paraTB were detected. The chance of obtaining a positive TB result by ELISA was seven times higher using the ELISA-paraTB method; hence, there is a significant risk of obtaining false-positive serological results for paraTB in M. bovis-infected cattle. However, the hypothesis that infection of M. bovis or prior TST performance may have boosted the host immune response and therefore increased the sensitivity of the paraTB-ELISA cannot be excluded.

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Development and diagnostic validation of an ELISA based on an antigenic mixture for the detection of bovine tuberculosis

Cited by 11 publications

References 21 publications

Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein

Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein

Review on Bovine Tuberculosis: An Emerging Disease Associated with Multidrug-Resistant Mycobacterium Species

The Risk of False-Positive Serological Results for Paratuberculosis in Mycobacterium bovis-Infected Cattle

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