Enzyme Linked Immunosorbent Assay (Elisa) for Detection of Clostridium Botulinum Type B Toxin

Kozaki, Shunji; Dufrenne, J.; Hagenaars, A. M.; Notermans, S.

doi:10.7883/yoken1952.32.199

“…Thorough validation of an ELISA for BoNT detection in different matrices is indispensable, because the extent of cross-reactivity between the antibodies used and the matrix components in an unknown sample is difficult to predict. Also when A, B, C, D, E, F, G 0.1-100 ng/mL 5 h-2 days cs, se, fo, fe Notermans et al (1978), Kozaki et al (1979), Notermans et al (1979), Lewis et al (1981), Lee and Yang (1982), Notermans et al (1982a, b), Dezfulian et al (1984), Michalik et al (1986, Thomas (1991), Doellgast et al (1993), Potter et al (1993), Doellgast et al (1994), Szílagyi et al (2000), Ferreira (2001), Ferreira et al (2001Ferreira et al ( , 2004, Poli et al (2002), Sharma et al (2006), Rajkovic et al (2012) ELISA (mAb)…”

Section: Laboratory Diagnostics Of Botulism: Conclusion and Perspectivesmentioning

confidence: 99%

Complexity of Botulinum Neurotoxins: Challenges for Detection Technology

Dorner

¹

,

Schulz

²

,

Kull

³

et al. 2012

Current Topics in Microbiology and Immunology

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The detection of botulinum neurotoxins (BoNT) is extremely challenging due to their high toxicity and the multiple BoNT variants. To date, seven serotypes with more than 30 subtypes have been described, and even more subtypes are expected to be discovered. The fact that the BoNT molecules are released as large complexes of different size and composition adds further complexity to the issue. Currently, in the diagnostics of botulism, the mouse bioassay (MBA) is still considered as gold standard for the detection of BoNT in complex sample materials. Over the years, different functional, immunological, and spectrometric assays or combinations thereof have been developed, supplemented by DNA-based assays for the detection of the organism. In this review, advantages and limitations of the current technologies will be discussed, highlighting some of the intricacies of real sample analysis.

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“…In order to quantify BoNT in a real sample, a pure BoNT standard is analyzed in parallel in defined concentrations. The use of classical immunoassays dates back to the late 1970s when the first sandwich ELISA systems for the detection of BoNT/A, B, and E were introduced (Notermans et al 1978(Notermans et al , 1979Kozaki et al 1979). They were soon complemented by ELISA for the detection of the other serotypes (Lewis et al 1981;Lee and Yang 1982;Notermans et al 1982b).…”

Section: Classical Sandwich Elisamentioning

confidence: 99%

Complexity of Botulinum Neurotoxins: Challenges for Detection Technology

Dorner

¹

,

Schulz

²

,

Kull

³

et al. 2012

Current Topics in Microbiology and Immunology

View full text Add to dashboard Cite

The detection of botulinum neurotoxins (BoNT) is extremely challenging due to their high toxicity and the multiple BoNT variants. To date, seven serotypes with more than 30 subtypes have been described, and even more subtypes are expected to be discovered. The fact that the BoNT molecules are released as large complexes of different size and composition adds further complexity to the issue. Currently, in the diagnostics of botulism, the mouse bioassay (MBA) is still considered as gold standard for the detection of BoNT in complex sample materials. Over the years, different functional, immunological, and spectrometric assays or combinations thereof have been developed, supplemented by DNA-based assays for the detection of the organism. In this review, advantages and limitations of the current technologies will be discussed, highlighting some of the intricacies of real sample analysis.

show abstract

“…In 1978 Notermans et al (50) used polystyrene tubes with polyclonal horse and rabbit antitoxin to detect type A toxin with a sensitivity of 50-1 00 LDso. In 1979 Kozaki et al (42) reported that a double sandwich ELISA technique improved the sensitivity from about 5,000 LDso to about 400 LDso for type B botulism toxin. When EDTA was added to toxic samples at a concentration of 5 mM just before incubation in the ELISA the ELISA extinction values were observed to rise twofold.…”

Section: Developmental Studiesmentioning

confidence: 99%