Modified toxin-binding inhibition (ToBI) test for epsilon antitoxin determination in serum of immunized rabbits

Santos, Eliane Macedo Sobrinho; Cangussu, Alex Sander Rodrigues; Brandi, Igor Viana; Sari, Roger Shoji; Almeida, Anna Christina de; Colen, Fernando; Quintilio, Wagner; Santos, Hércules Otacílio

doi:10.1016/j.vetimm.2010.07.007

Cited by 6 publications

(8 citation statements)

References 8 publications

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“…In South American and European countries, diagnoses of enteric diseases are performed by analyzing clinical signs, characterization of macroscopic and microscopic lesions, detection of the C. perfringens toxins type present in the intestinal contents of infected animals by ELISA and PCR, isolation of the etiological agent in bacteriological cultures, demonstration of toxins in clinical samples and in supernatant fluid from pure cultures, or by Seroneutralization of the intestinal filtrate combined with specific antitoxins (Sobrinho et al, 2010;Sobrinho et al, 2014;Santana et al, 2018;Ghoneim and Hamza, 2017). PCR is a basic molecular technique that can be employed to detect C. perfringens strains by in vitro Multiplex PCR tests, in which the corresponding toxin genes are detected.…”

Section: Diagnostic Methods For Enteric Diseasesmentioning

confidence: 99%

Pathologic findings on ruminant enteric clostridial diseases reveal specificities and differences among iota and iota-like toxins

Brito¹,

Alencar²,

Albuquerque³

et al. 2021

rcia

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The iota toxin (ITX) is a binary enterotoxin produced as a protoxin by Clostridium perfringens (C. perfringens) type E that is activated by proteolytic enzymes in the small intestine of infected animals. By depolymerization of the actin filaments, ITX causes cytoskeleton disorganization of cells promoting the increase of the cell permeability. Here, we conducted this review aiming to advance the understanding of enteric clostridial diseases caused by C. perfringens toxins and the specificity of ITX in the intestinal mucosa lesions. ITX consists of an enzymatic component (Ia) and a binding component (Ib). We screened the recently published histological findings of the ITX effects and its relationship with intestinal enteric diseases. Histologically, hemorrhagic necrosis and multifocal hemorrhage have been observed in the jejunum-ileum mucosa, the small intestine, and the abomasum. Although the diagnosis is still based on the presence of toxins in the intestinal contents and the clinical and/or histological history, it is important to develop novel enterotoxemic indicators capable of establishing precise methods for differentiate the actions of ITX and other toxins involved in the infectious process of C. perfringens type E.

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Section: Diagnostic Methods For Enteric Diseasesmentioning

confidence: 99%

Pathologic findings on ruminant enteric clostridial diseases reveal specificities and differences among iota and iota-like toxins

Brito¹,

Alencar²,

Albuquerque³

et al. 2021

rcia

View full text Add to dashboard Cite

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“…Clostridium perfringens Type D strains are responsible for enterotoxemia, which mainly affects sheep, but also occurs in goats and cattle, due to changes in intestinal microbiota after the appearance of symptoms. Infected animals may suffer sudden death with heavy economic losses (el Idrissi and Ward, 1992;Songer, 1996;Uzal and Songer, 2008;Sobrinho et al, 2010;Bokori-Brown et al, 2013). Epsilon toxin (Etx) is a pore-forming toxin and is produced by Clostridium perfringens strains belonging to toxin types B and D (Bokori-Brown et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Total combining power: Technique for the evaluation of the quality control process of clostridiosis vaccines

Brandi

Santos

Carvalho

et al. 2016

Journal of Microbiological Methods

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“…The toxin-antitoxin reaction is used to determine antibody levels in sera from immunized animals, and is used by official control agencies or vaccine manufacturers to assess the efficacy of clostridial vaccines and to certify their quality (Sobrinho et al, 2010). New methods for quantification of antigens are desired.…”

Section: Introductionmentioning

confidence: 99%

“…New methods for quantification of antigens are desired. This kit allows for the detection and quantification of antigens that are necessary for the vaccine formulation (Sobrinho et al, 2011). These tests, known as the toxin binding inhibition (ToBI) test and the Enzyme Linked Immuno Sorbent Assay (ELISA) aimed to evaluate the quality of veterinary vaccines in order to replace or minimize the procedures that use laboratory animals (Sobrinho et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…These tests, known as the toxin binding inhibition (ToBI) test and the Enzyme Linked Immuno Sorbent Assay (ELISA) aimed to evaluate the quality of veterinary vaccines in order to replace or minimize the procedures that use laboratory animals (Sobrinho et al, 2010). Other similar alternatives have been developed with the same purpose (Sobrinho et al, 2009 This study was triggered by the growing interest in the development of immunological techniques in vitro for the control of clostridial antigens, since the efficiency of the in vitro method in the detection of the toxin and toxoids, the advantages in terms of sensitivity and result quickness, as well as its relevancy in the routine of veterinary pharmaceutical industries, are well known Ward, 1992a, 1992b).…”

Section: Introductionmentioning

confidence: 99%

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ELISA and modified toxin-binding inhibition test for quality control of the clostridial vaccine processes

Santos¹,

Almeida

Brandi

et al. 2014

Arq. Bras. Med. Vet. Zootec.

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This study aimed to assess and standardize the ELISA and modified ToBI test in vitro methods in order to verify the potency of epsilon toxicoid in comparison with the in vivo TCP method. The following epsilon toxoids were used: NIBSC standard from batches 375/07, 532/08, 551/08, 373/07 and 378/07. These were evaluated using a TCP test, ELISA and ToBI tests. The results indicate that the correlation ratio between the dilutions of standard NIBSC toxicoid and absorbance values of 89.44% obtained with the ELISA method support the use of the curve to evaluate epsilon toxoids. However, it was observed that the absorbance values were similar for all toxoids, thus presenting no significant difference between higher and lower concentration toxoids. For the ToBI test, the correlation ratio of 96.76, obtained in the curve pattern, demonstrates the effectiveness of the curve to be used in the epsilon toxoid evaluation. The correlation ratio between the titration degrees of toxoids obtained through TCP and ToBI tests was higher than 90%. It is concluded that the type of ELISA test used does present discriminative power for toxoids with different concentrations, which does not support the use of this technique for such a purpose. The ToBI test can be used as a screening method for it is sensitive and effective to detect epsilon toxicoid produced by C. perfringens type D.

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Modified toxin-binding inhibition (ToBI) test for epsilon antitoxin determination in serum of immunized rabbits

Cited by 6 publications

References 8 publications

Pathologic findings on ruminant enteric clostridial diseases reveal specificities and differences among iota and iota-like toxins

Pathologic findings on ruminant enteric clostridial diseases reveal specificities and differences among iota and iota-like toxins

Total combining power: Technique for the evaluation of the quality control process of clostridiosis vaccines

ELISA and modified toxin-binding inhibition test for quality control of the clostridial vaccine processes

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