Quantitative anti-PA IgG ELISA; assessment and comparability with the anthrax toxin neutralization assay in goats

Ndumnego, Okechukwu Chinazo; Crafford, Jan Ernst; Beyer, Wolfgang; Heerden, Henriëtte van

doi:10.1186/1746-6148-9-265

Cited by 11 publications

(25 citation statements)

References 35 publications

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“…B. anthracis spores have the capacity to contaminate a given area for a long time, because of their intrinsic ability to survive different environmental conditions and chemical disinfectants [ 2 , 8 ]. Anthrax spores have been isolated after 60 years from contaminated sites [ 8 , 9 ]. B. anthracis has been recovered from animal bones estimated to be over 200 years old in the Kruger National Park in South Africa [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

The efficacy and safety of nine South African medicinal plants in controlling Bacillus anthracis Sterne vaccine strain

Elisha

Dzoyem

Botha

et al. 2015

BMC Complement Altern Med

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BackgroundAnthrax is a zoonotic disease caused by Bacillus anthracis, a Gram-positive spore-forming bacterium. The presence of the bacteria and the toxins in the blood of infected hosts trigger a cascade of pathological events leading to death. Nine medicinal plants with good activities against other bacteria were selected to determine their in vitro antibacterial activity against Bacillus anthracis Sterne strain. The cytotoxicity of the extracts on Vero kidney cells was also determined.ResultsThe minimum inhibitory concentration (MIC) values of the extracts against Bacillus anthracis Sterne strain ranged from 0.02 to 0.31 mg/ml. Excellent MIC values were observed for the following plant species: Maesa lanceolata (0.02 mg/ml), Bolusanthus speciosus, Hypericum roeperianum, Morus mesozygia (0.04 mg/ml) and Pittosporum viridiflorum (0.08 mg/ml). The total antibacterial activity of the extracts ranged from 92 to 5562 ml/g. Total activity presents the volume to which the extract from 1 g of plant material can be diluted and still inhibit microbial growth. Maesa lanceolata and Hypericum roeperianum had the highest total activity with values of 5562 and 2999 ml/g respectively. The extracts of Calpurnia aurea had the lowest total activity (92 ml/g). The cytotoxicity determined on Vero cells indicated that most of the extracts were relatively non-toxic compared to doxorubicin (LC50 8.3 ± 1.76 μg/ml), except for the extracts of Maesa lanceolata, Elaeodendron croceum and Calpurnia aurea with LC50 values at 2.38 ± 0.25, 5.20 ± 0.24 and 13 ± 2.26 μg/ml respectively. The selectivity index (SI) ranged from 0.02 to 1.66. Hypericum roeperianum had the best selectivity index, (SI = 1.66) and Elaeodendron croceum had lowest value (SI = 0.02).ConclusionsThe crude acetone extracts of the selected plant species had promising antibacterial activity against Bacillus anthracis. Maesa lanceolata extracts could be useful as a disinfectant and Hypericum roeperianum could be useful to protect animals based on its high total activity and selectivity index. Further investigation of these plant extracts may lead to the development of new therapeutic agents to protect humans or animals against anthrax.

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Section: Introductionmentioning

confidence: 99%

The efficacy and safety of nine South African medicinal plants in controlling Bacillus anthracis Sterne vaccine strain

Elisha

Dzoyem

Botha

et al. 2015

BMC Complement Altern Med

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“…Sera collected at weeks 0, 4 and 17 were analysed for PA-specific antibody using the ELISA as previously described by Hahn et al [ 15 ] and Pombo et al [ 16 ] with some modifications as described by Ndumnego et al [ 17 ]. Endpoint titres of individual serum were defined as the reciprocal of the highest serum dilution giving an optical density of 0.1.…”

Section: Methodsmentioning

confidence: 99%

“…An in vitro toxin neutralization assay (TNA) was performed using a MTT [3-(4.5-dimethylthiazol-2-yl)–2.5-diphenyltetrazolium bromide] in a colorimetric cell viability assay with the J774A.1 macrophage cell line as previously described by Hering et al [ 18 ] with slight modifications by Ndumnego et al [ 17 ]. Briefly, serial diluted goat serum was incubated with PA and LF (List Biological Laboratories, USA) at concentrations of 500 and 400 ng/mL respectively before addition to overnight incubated macrophage cells.…”

Section: Methodsmentioning

confidence: 99%

Use of the mice passive protection test to evaluate the humoral response in goats vaccinated with Sterne 34F2 live spore vaccine

Phaswana

Ndumnego

Koehler

et al. 2017

Vet Res

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The Sterne live spore vaccine (34F2) is the most widely used veterinary vaccine against anthrax in animals. Antibody responses to several antigens of Bacillus anthracis have been described with a large focus on those against protective antigen (PA). The focus of this study was to evaluate the protective humoral immune response induced by the live spore anthrax vaccine in goats. Boer goats vaccinated twice (week 0 and week 12) with the Sterne live spore vaccine and naive goats were used to monitor the anti-PA and toxin neutralizing antibodies at week 4 and week 17 (after the second vaccine dose) post vaccination. A/J mice were passively immunized with different dilutions of sera from immune and naive goats and then challenged with spores of B. anthracis strain 34F2 to determine the protective capacity of the goat sera. The goat anti-PA ELISA titres indicated significant sero-conversion at week 17 after the second doses of vaccine (p = 0.009). Mice receiving undiluted sera from goats given two doses of vaccine (twice immunized) showed the highest protection (86%) with only 20% of mice receiving 1:1000 diluted sera surviving lethal challenge. The in vitro toxin neutralization assay (TNA) titres correlated to protection of passively immunized A/J mice against lethal infection with the vaccine strain Sterne 34F2 spores using immune goat sera up to a 1:10 dilution (rs ≥ 0.522, p = 0.046). This study suggests that the passive mouse protection model could be potentially used to evaluate the protective immune response in livestock animals vaccinated with the current live vaccine and new vaccines.

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“…The reaction was stopped by adding 50µl of HCL 5.8% and absorbance was read in wavelength 450 nm against 630 nm second filters in an ELISA reader (Stat Fax. USA) and ELISA value was obtained (Ndumnego OC et al, 2013). All washing steps were carried out with micro plate washer (STAT FAX, USA).…”

Section: Elisa For Anti-pa4 and Anti-lf Antibody Measurementmentioning

confidence: 99%

“…In the follow-up step of ELISA, reactions were stopped by adding 50µl of Hcl5.8%, and the optical density (OD) at 450 nm and 630 was determined and ELISA value was obtained. (Ndumnego et al, 2013;Mitic et al, 2016).…”

Section: Competitive Inhibition Elisamentioning

confidence: 99%

Development of Enzyme Linked Immunosorbent Assay for humoral immuneresponse and infection monitoring of anthrax

Nouri

Razzaz

Taghdiri

et al. 2020

J Hellenic Vet Med Soc

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Immune assays were taken into consideration to diagnose and quantify metabolites such as antigen and antibody. Enzyme-Linked Immunosorbent Assays (ELISAs), which are used to detect antigens and antibodies, generated several periods of infectious and vaccination conditions. There is an extensive range of commercial infectious disease ELISA kits useful for the detection of human and animal IgG, IgA, IgM antibodies and microorganism antigens. Anthrax is one of the serious infectious diseases caused by rod-shaped, gram-positive bacteria known as Bacillus anthracis. Subunit or attenuated vaccines applied against anthrax disease increase the antibody against the Protective Antigen (PA) which has a critical role as a toxin of B. anthracis. Herein, the ELISA was developed using PA domain 4 and anthrax Lethal Factor to detect IgG antibody in serum. Besides, the level of anti-LF antibodies were determined as a complementary test to measure variance in antibody titers associated with vaccination or infection that leads to detection of anthrax in livestock. The results show that we developed high-quality ELISA kit that can be used to test immunogenicity of vaccines and infections in mice. We tried to develop the Anti- PA4 ELISA kit and conduct the validation studies to evaluate the fluctuation level of the antibody in the anthrax vaccine and distinction between disease and vaccination in mice.

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Quantitative anti-PA IgG ELISA; assessment and comparability with the anthrax toxin neutralization assay in goats

Cited by 11 publications

References 35 publications

The efficacy and safety of nine South African medicinal plants in controlling Bacillus anthracis Sterne vaccine strain

The efficacy and safety of nine South African medicinal plants in controlling Bacillus anthracis Sterne vaccine strain

Use of the mice passive protection test to evaluate the humoral response in goats vaccinated with Sterne 34F2 live spore vaccine

Development of Enzyme Linked Immunosorbent Assay for humoral immuneresponse and infection monitoring of anthrax

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