Production and Characterization of Monoclonal Antibodies Specific for Mycobacterium bovis

Wood, P.R.; Ripper, Jan; Radford, Anthony J.; Bundesen, Peter; Rylatt, Dennis B.; Cottis, L.E.; John, Manohar; Plackett, P.

doi:10.1099/00221287-134-9-2599

Cited by 27 publications

(30 citation statements)

References 15 publications

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“…1) A second area, containing epitopes for SB5,6, 7 and 8 was mapped towards the carboxyl terminal of the molecule. Competition analysis of 10 SB MoAb showed that they could be grouped into three sets (23). SB5, 6, 7 and 8 competed with each other indicating that they bound to the same or neighbouring epitopes.…”

Section: Discussionmentioning

confidence: 99%

“…The recombinant GST fusion antigens were tested in an EIA for reactivity with nine MoAb which recognize at least three separate epitopes on MPB70 (Table 3) (23). The controls were the same as in the T cell assays.…”

Section: Analysis Of B Cell Epitopes By Eiamentioning

confidence: 99%

“…The filters were immunoblotted with anti-MPB70 MoAb, SBIO (23). Total cell protein extracts made from reactive clones were run on an SDS-PAGE gel and then transferred on to nitrocellulose and probed with SB 10 to confirm reactivity with the MoAb and to estimate the size of the recombinant MPB70.…”

Section: Construction Of Deletion Mutants Of Mpb70mentioning

confidence: 99%

“…The EIA method used was adapted from Wood et al (23). Wells of polystyrene microtitre trays were coated overnight at 4°C with 1 ng of antigen diluted in PBS.…”

Section: Eia Of Recombinant Antigens Using Monoclonal Antibodiesmentioning

confidence: 99%

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Mapping of the T and B cell epitopes of the Mycobacterium bovis protein, MPB70

Billman‐Jacobe

Radford

Rothel

et al. 1990

Immunology & Cell Biology

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Summary A clone coding for the entire gene for the Mycobacterium bovis protein antigen MPB70 was used to produce a series of overlapping subclones by making a series of deletions from the 3' end of the gene. The subclones expressed incomplete MPB70 proteins as fusions with glutathione-Stransferase. The insert DNA was sequenced to determine the extent of the deletion and the proteins expressed by the clones were examined for the presence of T cell and B cell epitopes. T cell epitopes were mapped by measuring the ability of recombinant antigens to stimulate gamma interferon (7-IFN) production in a whole blood culture system. y-IFN production was measured using a sandwich enzyme immunoasssay specific for bovine y-IFN. B cell epitopes were mapped with a series of anti-MPB70 monoclonal antibodies using an indirect enzyme immunoassay.

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Section: Discussionmentioning

confidence: 99%

Section: Analysis Of B Cell Epitopes By Eiamentioning

confidence: 99%

Section: Construction Of Deletion Mutants Of Mpb70mentioning

confidence: 99%

“…The EIA method used was adapted from Wood et al (23). Wells of polystyrene microtitre trays were coated overnight at 4°C with 1 ng of antigen diluted in PBS.…”

Section: Eia Of Recombinant Antigens Using Monoclonal Antibodiesmentioning

confidence: 99%

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Mapping of the T and B cell epitopes of the Mycobacterium bovis protein, MPB70

Billman‐Jacobe

Radford

Rothel

et al. 1990

Immunology & Cell Biology

Self Cite

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“…and TB1 R (5?-AGC ACG CTG TCA ATC ATG TA-3?) amplify a 372-bp region of the gene responsible for the production of MPB 70, a secretory protein considered to be specific for M. tuberculosis complex (Wood et al 1988;Cousins et al 1991).…”

Section: Laboratory Proceduresmentioning

confidence: 99%

Comparison of different methods for diagnosis of bovine tuberculosis from tuberculin- or interferon-γ-reacting cattle in Spain

Llamazares¹,

Gutiérrez-Martín²,

Martín³

et al. 1999

Journal of Applied Microbiology

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GONZÁ LEZ LLAMAZARES, O.R., GUTIÉ RREZ MARTÍN, C.B., ARANAZ MARTÍN, A., LIÉ BANA CRIADO, E., DOMÍNGUEZ RODRÍGUEZ, L. AND RODRÍG U EZ FE R RI , E . F. 1999. Of 1479 cattle from herds in Northwestern Spain previously diagnosed as tuberculosis (TB) positive, 218 animals which gave a positive tuberculin or interferon-g reaction were examined at the slaughterhouse. Medial retropharyngeal and caudal mediastinal lymph nodes, and any tissues containing lesions suspected to be tuberculous, were removed and submitted to the laboratory. Three techniques for diagnosis of TB were used: post mortem examination (PME), smear staining by means of auramine O method (AOM), and culture isolation in Coletsos and Lowenstein-Jensen media followed by confirmation of M. tuberculosis complex organisms using PCR (CIM-PCR). Only 123 (29·9%) of the 412 samples collected showed typical tuberculous lesions. Confirmed M. tuberculosis complex organisms were isolated in 144 cases, 114 of which were from tissues showing lesions (success rate of 92·8%). Smears were found positive in 113 cases, 96 of which came from lesions suspected to be tuberculous (success rate of 78·0%). The sensitivities of CIM-PCR compared with those of PME and AOM were 92·7% and 85·7%, respectively. Statistical analysis revealed that PME and AOM are good indicators of the presence of M. tuberculosis complex organisms in tuberculinor interferon-g reacting cattle.

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