Development and evaluation of a multi-antigen serological assay for the intra-vitam diagnosis of Tuberculosis caused by Mycobacterium bovis in pigs

Ippolito, Dorotea; Boniotti, M. B.; Fiasconaro, M.; Fontana, Stefania; Boifava, Mery; Ciarello, Flavia Pruiti; Amato, Benedetta; Pacciarini, Maria Lodovica; Presti, Vincenzo Di Marco Lo

doi:10.1016/j.jim.2022.113234

Cited by 3 publications

(5 citation statements)

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“…The multiplexed Luminex platform is used for many serological applications in veterinary medicine [ 54 – 57 ]. However, few studies have involved the use of this technology for the serological diagnosis of bTB [ 17 , 24 ]. In the present study, a panel of antigens including four major known immunogenic proteins of M .…”

Section: Discussionmentioning

confidence: 99%

“…The multiplexed Luminex platform is used for many serological applications in veterinary medicine [54][55][56][57]. However, few studies have involved the use of this technology for the serological diagnosis of bTB [17,24]. In the present study, a panel of antigens including four major known immunogenic proteins of M. bovis (MPB70, MPB83, the MPB70/83 fusion protein, and the ESAT6/CFP10 heterodimer), the PE25/PPE41 fusion protein, and four other proteins (Mb1961c, Mb1301c, Mb1403, Mb3871, and Mb0592) was selected and used to develop and evaluate a multiplexed serological test based on Luminex technology for the detection of bTB in cattle.…”

Section: Discussionmentioning

confidence: 99%

“…In two recent studies, Luminex assays employing immunodominant M . bovis antigens (MPB70, MPB83, ESAT6, and CFP10) were used for bTB diagnosis in cattle [ 17 ] and pigs [ 24 ].…”

Section: Introductionmentioning

confidence: 99%

“…After serum incubation, antigen-antibody reactions are detected using a dual laser system based on the principle of flow cytometry [22,23]. In two recent studies, Luminex assays employing immunodominant M. bovis antigens (MPB70, MPB83, ESAT6, and CFP10) were used for bTB diagnosis in cattle [17] and pigs [24].…”

Section: Introductionmentioning

confidence: 99%

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Identification of New Mycobacterium bovis antigens and development of a multiplexed serological bead-immunoassay for the diagnosis of bovine tuberculosis in cattle

Moens,

Filée,

Boes

et al. 2023

PLoS ONE

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Serological assays for bovine tuberculosis diagnosis require the use of multiple Mycobacterium bovis specific antigens to ensure the detection of infected animals. In the present study, identification and selection process of antigens, based on data from published proteomic studies and involving the use of bioinformatics tools and an immuno-screening step, was firstly performed for identifying novel antigens that elicit an antibody response in M. bovis infection. Based on this approach, a panel of 10 M. bovis antigens [with known relevance (MPB70, MPB83, MPB70/83, and ESAT6/CFP10) and novel (Mb1961c, Mb1301c, Mb3871, Mb1403, Mb0592, and PE25/PPE41)] were constructed and thenused to develop a new multiplexed serological assay based on Luminex technology. The performance of the Luminex-bTB immunoassay was evaluated using sera from cattle with known tuberculosis status. Among the proteins whose ability to detect bovine tuberculosis was evaluated for the first time, PE25/PPE41 and Mb1403, but not Mb3871, showed good detection capacity. Following multiple antigen combination, the final Luminex-bTB immunoassay included seven antigens (MPB70, MPB83, MPB70/83, ESAT6/CFP10, PE25/PPE41, Mb1403, and Mb0592) and showed better global performance than the immunoassay using the four usual antigens (MPB70, MPB70/83, MPB83 and ESAT6/CFP10). The specificity and sensitivity values were, respectively, of 97.6% and 42.8% when the cut-off of two-positive antigens was used to classify samples as positive. With the use of the more-restrictive criterion of three-positive antigens, the specificity increased to 99.2% but the sensitivity decreased to 23.9%. The analysis of antigen profiles generated with the Luminex-bTB immunoassay showed that mainly serodominant proteins were recognized in samples from infected cattle. The detection of Mb1961c and Mb1301c appeared to be associated with presumed false-positive results. Moreover, sera from cattle originating from bTB-outbreaks but having inconclusive or negative skin test results were identified as positive by the Luminex-bTB immunoassay and showed an antigen pattern associated with M. bovis infection. The Luminex-bTB immunoassay including seven antigens may be useful as adjunct test for the detection of M. bovis–infected herds, and different cut-offs could be applied according to the bovine tuberculosis epidemiological context.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…In two recent studies, Luminex assays employing immunodominant M . bovis antigens (MPB70, MPB83, ESAT6, and CFP10) were used for bTB diagnosis in cattle [ 17 ] and pigs [ 24 ].…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of New Mycobacterium bovis antigens and development of a multiplexed serological bead-immunoassay for the diagnosis of bovine tuberculosis in cattle

Moens,

Filée,

Boes

et al. 2023

PLoS ONE

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“…In particular, serological tests for determining antibodies to MBT antigens MPT63, MPT64 [16], and MPT83 [17,18] have a good prospect for widespread use.…”

mentioning

confidence: 99%

Detection of Specific Antibodies Using Mpt83-Mpt63 Fusion Protein in Patients With Destructive Pulmonary Tuberculosis

Rekalova

Panasiukova²,

Pohrebna³

et al. 2022

PSSS MS

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Introduction. The development of eff ective test systems for tuberculosis (TB) diagnosis and the study of factors infl uencing their sensitivity and specifi city constitute urgent tasks given the unstable epidemiological situation with TB in Ukraine and the world. The study aimed to determine the clinical and laboratory features of pulmonary TB that may aff ect the sensitivity of IgG TB antibody detection in human serum by the immuno-enzyme test system "IB-Chem Anti-Mycobacterium tuberculosis", which was developed at the Palladin Institute of Biochemistry of the NAS of Ukraine and uses recombinant fusion protein MPT83-MPT63 as an antigen. Methods. Sera of 62 patients with chemoresistant pulmonary TB were tested for TB antibodies using an indirect enzyme-linked immunosorbent assay. Dependence of the obtained results on clinical symptoms and signs of pulmonary TB, the frequency of diff erent types of antibiotic resistance, cellular and biochemical parameters of blood were analysed. Results. The sensitivity of the immuno-enzyme test system "IB-Chem Anti-Mycobacterium tuberculosis" for detecting human IgG TB antibodies using recombinant fusion protein MPT83-MPT63 was found to constitute 71.0%; it reached 81.3% in patients with destructive pulmonary TB. Conclusion. The developed test system should primarily be used for the TB diagnosis in patients with destructive changes in the lungs where the diagnostic effi ciency of this test system reaches high indicators.

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Evidence, Challenges, and Knowledge Gaps Regarding Latent Tuberculosis in Animals

et al. 2022

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Mycobacterium bovis and other Mycobacterium tuberculosis complex (MTBC) pathogens that cause domestic animal and wildlife tuberculosis have received considerably less attention than M. tuberculosis, the primary cause of human tuberculosis (TB). Human TB studies have shown that different stages of infection can exist, driven by host–pathogen interactions. This results in the emergence of heterogeneous subpopulations of mycobacteria in different phenotypic states, which range from actively replicating (AR) cells to viable but slowly or non-replicating (VBNR), viable but non-culturable (VBNC), and dormant mycobacteria. The VBNR, VBNC, and dormant subpopulations are believed to underlie latent tuberculosis (LTB) in humans; however, it is unclear if a similar phenomenon could be happening in animals. This review discusses the evidence, challenges, and knowledge gaps regarding LTB in animals, and possible host–pathogen differences in the MTBC strains M. tuberculosis and M. bovis during infection. We further consider models that might be adapted from human TB research to investigate how the different phenotypic states of bacteria could influence TB stages in animals. In addition, we explore potential host biomarkers and mycobacterial changes in the DosR regulon, transcriptional sigma factors, and resuscitation-promoting factors that may influence the development of LTB.

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Development and evaluation of a multi-antigen serological assay for the intra-vitam diagnosis of Tuberculosis caused by Mycobacterium bovis in pigs

Cited by 3 publications

References 32 publications

Identification of New Mycobacterium bovis antigens and development of a multiplexed serological bead-immunoassay for the diagnosis of bovine tuberculosis in cattle

Identification of New Mycobacterium bovis antigens and development of a multiplexed serological bead-immunoassay for the diagnosis of bovine tuberculosis in cattle

Detection of Specific Antibodies Using Mpt83-Mpt63 Fusion Protein in Patients With Destructive Pulmonary Tuberculosis

Evidence, Challenges, and Knowledge Gaps Regarding Latent Tuberculosis in Animals

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