Development of sensitive colorimetric capture elisas for Clostridium botulinum neurotoxin serotypes A and B

Sensing the Deadliest Toxin: Technologies for Botulinum Neurotoxin Detection

“…The experiment usually takes 5-6 hours to complete, and is significantly faster than mouse lethality assay. ELISAs with an experimental setup analogous to this were reported to have a wide range of sensitivities, depending on the specific antibody and reporter system used, and have been employed for detection of all BoNT serotypes [48,49,50,51]. …”

“…ELISA methodologies have been successfully employed in the detection and quantification of purified botulinum toxin [49,51,59], in C. botulinum cultures that produce the toxin [60,61,62], in an extensive variety of food samples [52,54,63,64,65] (both contaminated food and food artificially spiked with the toxin), and in clinical samples such as serum [49] and feces [66]. Some foods tend to interfere with ELISAs and decrease their sensitivity; therefore, results should be confirmed by mouse lethality assay.…”

Rapid detection of Clostridium botulinum toxins A, B, E, and F in clinical samples, selected food matrices, and buffer using paramagnetic bead-based electrochemiluminescence detection

“…One problem with such assays is the susceptibility of the assay substrates to nonrelated proteases and the consequent risk of false positives. A variety of antibody-based assays that are both sensitive and serotype specific have been proposed [5][6][7][8]. A drawback with all immunoassays is their inability to distinguish biologically active toxin or toxin fragments from biologically inactive ones.…”

Complexity of Botulinum Neurotoxins: Challenges for Detection Technology

“…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)…”

“…Results were routinely obtained within 4-6 h, a clear advancement in comparison to the MBA. However, when using polyclonal antibodies (pAb) generated against purified BoNT or BoNT complexes, the ELISAs were clearly less sensitive than the MBA with detection limits usually down to a few ng/mL (Thomas 1991;Doellgast et al 1993Doellgast et al , 1994Szílagyi et al 2000;Ferreira 2001;Ferreira et al 2004;Sharma et al 2006). Later, in a collaborative effort, sandwich ELISAs based on pAb against BoNT/A, B, E, and F have been validated and compared to the MBA, resulting in assay sensitivities of 0.1-1 ng/ mL.…”