Advanced search
1 file | 564.96 KB Add to list

Detection of Clostridium botulinum neurotoxins A and B in milk by ELISA and immuno-PCR at higher sensitivity than mouse bio-assay

(2012) FOOD ANALYTICAL METHODS. 5(3). p.319-326
Author
Organization
Abstract
Using commercially available antibodies and toxoids as templates, an ELISA, immuno-quantitative PCR (iqPCR), and multiplex immuno-PCR (iPCR) were developed for detection of Clostridium botulinum neurotoxins A and B. The obtained sensitivities for ELISA ranged from 1 ng/ml in PBS + 1% BSA to 15 and 10 ng/ml in skimmed milk for botulinum neurotoxins (BoNT)/A and BoNT/B, respectively. In semi-fat milk, the limit of detection (LOD) for both toxoids was 30 ng/ml. Quantitative immuno-PCR (iqPCR) had an LOD of 4.5-9 pg/reaction for BoNT/A in both PBS and semi-fat milk, while this was 18.5-37 pg/reaction for BoNT/B in PBS + 1% BSA and semi-fat milk, respectively. The sensitivities of ELISA and iqPCR were improved to 0.5 ng/ml and 3.75 pg/ml (0.2 pg/reaction) in semi-fat milk, respectively, when toxoid of BoNT/A was substituted with actual toxin. Multiplex iPCR with both toxoids run in the same reaction was able to distinguish presence/absence of tested BoNT/A and BoNT/B at 25 pg/reaction. The tested system offers a realistic alternative with much better sensitivity to the standard mouse assay.
Keywords
Clostridium botulinum, Neurotoxin, ELISA, Immuno-PCR, Detection, Food, Milk, POLYMERASE-CHAIN-REACTION, RAPID DETECTION, TOXIN, ANTIBODIES, FOODS, ASSAY, IMMUNOGENICITY, QUANTIFICATION, DIAGNOSTICS, BIOSENSOR

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 564.96 KB

Citation

Please use this url to cite or link to this publication:

MLA
Rajkovic, Andreja, Benaissa El Moualij, Youssef Fikri, et al. “Detection of Clostridium Botulinum Neurotoxins A and B in Milk by ELISA and immuno-PCR at Higher Sensitivity Than Mouse Bio-assay.” FOOD ANALYTICAL METHODS 5.3 (2012): 319–326. Print.
APA
Rajkovic, A., El Moualij, B., Fikri, Y., Dierick, K., Zorzi, W., Heinen, E., Üner, A., et al. (2012). Detection of Clostridium botulinum neurotoxins A and B in milk by ELISA and immuno-PCR at higher sensitivity than mouse bio-assay. FOOD ANALYTICAL METHODS, 5(3), 319–326.
Chicago author-date
Rajkovic, Andreja, Benaissa El Moualij, Youssef Fikri, Katelijne Dierick, Willy Zorzi, Ernst Heinen, Ahu Üner, and Mieke Uyttendaele. 2012. “Detection of Clostridium Botulinum Neurotoxins A and B in Milk by ELISA and immuno-PCR at Higher Sensitivity Than Mouse Bio-assay.” Food Analytical Methods 5 (3): 319–326.
Chicago author-date (all authors)
Rajkovic, Andreja, Benaissa El Moualij, Youssef Fikri, Katelijne Dierick, Willy Zorzi, Ernst Heinen, Ahu Üner, and Mieke Uyttendaele. 2012. “Detection of Clostridium Botulinum Neurotoxins A and B in Milk by ELISA and immuno-PCR at Higher Sensitivity Than Mouse Bio-assay.” Food Analytical Methods 5 (3): 319–326.
Vancouver
1.
Rajkovic A, El Moualij B, Fikri Y, Dierick K, Zorzi W, Heinen E, et al. Detection of Clostridium botulinum neurotoxins A and B in milk by ELISA and immuno-PCR at higher sensitivity than mouse bio-assay. FOOD ANALYTICAL METHODS. 2012;5(3):319–26.
IEEE
[1]
A. Rajkovic et al., “Detection of Clostridium botulinum neurotoxins A and B in milk by ELISA and immuno-PCR at higher sensitivity than mouse bio-assay,” FOOD ANALYTICAL METHODS, vol. 5, no. 3, pp. 319–326, 2012.
@article{2102757,
  abstract     = {Using commercially available antibodies and toxoids as templates, an ELISA, immuno-quantitative PCR (iqPCR), and multiplex immuno-PCR (iPCR) were developed for detection of Clostridium botulinum neurotoxins A and B. The obtained sensitivities for ELISA ranged from 1 ng/ml in PBS + 1% BSA to 15 and 10 ng/ml in skimmed milk for botulinum neurotoxins (BoNT)/A and BoNT/B, respectively. In semi-fat milk, the limit of detection (LOD) for both toxoids was 30 ng/ml. Quantitative immuno-PCR (iqPCR) had an LOD of 4.5-9 pg/reaction for BoNT/A in both PBS and semi-fat milk, while this was 18.5-37 pg/reaction for BoNT/B in PBS + 1% BSA and semi-fat milk, respectively. The sensitivities of ELISA and iqPCR were improved to 0.5 ng/ml and 3.75 pg/ml (0.2 pg/reaction) in semi-fat milk, respectively, when toxoid of BoNT/A was substituted with actual toxin. Multiplex iPCR with both toxoids run in the same reaction was able to distinguish presence/absence of tested BoNT/A and BoNT/B at 25 pg/reaction. The tested system offers a realistic alternative with much better sensitivity to the standard mouse assay.},
  author       = {Rajkovic, Andreja and El Moualij, Benaissa and Fikri, Youssef and Dierick, Katelijne and Zorzi, Willy and Heinen, Ernst and Üner, Ahu and Uyttendaele, Mieke},
  issn         = {1936-9751},
  journal      = {FOOD ANALYTICAL METHODS},
  keywords     = {Clostridium botulinum,Neurotoxin,ELISA,Immuno-PCR,Detection,Food,Milk,POLYMERASE-CHAIN-REACTION,RAPID DETECTION,TOXIN,ANTIBODIES,FOODS,ASSAY,IMMUNOGENICITY,QUANTIFICATION,DIAGNOSTICS,BIOSENSOR},
  language     = {eng},
  number       = {3},
  pages        = {319--326},
  title        = {Detection of Clostridium botulinum neurotoxins A and B in milk by ELISA and immuno-PCR at higher sensitivity than mouse bio-assay},
  url          = {http://dx.doi.org/10.1007/s12161-011-9300-7},
  volume       = {5},
  year         = {2012},
}

Altmetric
View in Altmetric
Web of Science
Times cited: