Skip to main content

Advertisement

Log in

Comparison of immunostrips with mouse bioassay and bacterial culture in detecting botulinum toxins in bottles from suspected Taiwan high-speed rail bomber

  • Original Article
  • Published:
Forensic Toxicology Aims and scope Submit manuscript

Abstract

In April 2013, a bomb-threat incident occurred on the Taiwan high-speed railway. Two suspects claimed to have placed explosive devices on a high-speed train and outside a lawmaker’s office. Afterward, the Criminal Investigation Bureau also found several bottles in the main suspect’s apartment labeled “botulinum toxin.” The Institute of Preventive Medicine was entrusted by the Institute of Forensic Medicine, Ministry of Justice, to verify whether the bottles contained botulinum neurotoxin (BoNT). Three different analyses, including rapid detection strips, toxin neutralization tests, and bacterial culture, were processed. The results provided conclusive evidence that botulinum neurotoxin type A (BoNT/A) was present in the samples. This study demonstrated that the BoNT/A strip assay is an excellent method for BoNT/A detection, and has potential use as an early warning tool in screening of food products for botulinum toxins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Peck MW (2009) Biology and genomic analysis of Clostridium botulinum. Adv Microb Physiol 55:183–265

    Article  CAS  PubMed  Google Scholar 

  2. Bruggemann H, Woller A, Mazuet C, Popoff MR (2011) Clostridium botulinum. Genomes of foodborne and waterborne pathogens. In: Fratamico P, Liu Y, Kathariou S (eds). Washington pp 185–212

  3. Hill KK, Smith TJ, Helma CH, Ticknor LO, Foley BT (2007) Genetic diversity among botulinum neurotoxin-producing clostridial strains. J Bacteriol 189:818–832

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Cherington M (2004) Botulism: update and review. Semin Neurol 24:155–163

    Article  PubMed  Google Scholar 

  5. Johnson EA (1999) Clostridial toxins as therapeutic agents: benefits of nature’s most toxic proteins. Annu Rev Microbiol 53:551–575

    Article  CAS  PubMed  Google Scholar 

  6. Middlebrook JL (1989) Cell surface receptors for protein toxins. In: Simpson LL (ed) Botulinum neurotoxin and tetanus toxin. Academic, New York, pp 95–119

    Chapter  Google Scholar 

  7. Niemann H (1991) Molecular biology of the clostridial neurotoxins. In: Alouf JE, Freer JH (eds) A sourcebook on bacterial protein toxins. Academic, London, pp 303–348

    Google Scholar 

  8. Ting PT, Freiman A (2004) The story of Clostridium botulinum: from food poisoning to Botox. Clin Med 4:258–261

    Article  PubMed  Google Scholar 

  9. Walt DR, Franz DR (2000) Biological warfare detection. Anal Chem 72:738–746

    Article  Google Scholar 

  10. Iura K, Tsuge K, Seto Y, Sato A (2004) Detection of proteinous toxins using the Bio Threat Alert system. Jpn J Forensic Toxicol 22:13–16

    CAS  Google Scholar 

  11. Tsuge K, Ohsawa I, Matsushita K, Sekiguchi H, Seto Y, Sato A (2005) Detection of proteinous toxins using the Bio Threat Alert System. 2. Response to high levels of toxins and interference by colored sample matrix. Jpn J Forensic Toxicol 23:18–20

    CAS  Google Scholar 

  12. Sano Y, Yamashiro S, Komano A, Maruko H, Sekiguchi H, Takayama Y, Sekioka R, Tsuge K, Ohsawa I, Kanamori-Kataoka M, Seto Y, Satoh A (2007) Detection of proteinous toxins using the Bio-Threat Alert system, part 3: effects of heat pretreatment and interfering substances. Forensic Toxicol 25:76–79

    Article  CAS  Google Scholar 

  13. Yamashiro S, Sano Y, Komano A, Maruko H, Sekiguchi H, Takayama Y, Sekioka R, Tsuge K, Ohsawa I, Kanamori-Kataoka M, Seto Y, Satoh A (2007) Detection of proteinous toxins using the Bio-Threat Alert system, part 4. Differences in test strip manufacturing lot and toxin subtype detectability. Forensic Toxicol 25:80–83

    Article  CAS  Google Scholar 

  14. Franz DR, Zajtchuk R (2000) Biological terrorism: understanding the threat, preparation, and medical response. Dis Mon 46:125–190

    Article  CAS  PubMed  Google Scholar 

  15. Hansen JE (1999) Viruses, bacteria and toxins as biological warfare (Danish). Ugeskr Laeger 161:772–775

    CAS  PubMed  Google Scholar 

  16. Bossi P, Bricaire F (2003) Botulism toxin, bioterrorist weapon. Presse Med 32:463–465

    PubMed  Google Scholar 

  17. Arnon SS, Schechter R, Inglesby TV, Henderson DA, Bartlett JG, Ascher MS, Eitzen E, Fine AD, Hauer J, Layton M, Lillibridge S, Osterholm MT, O’Toole T, Parker G, Perl TM, Russell PK, Swerdlow DL, Tonat K (2001) Botulinum toxin as a biological weapon: medical and public health management. JAMA 285:1059–1070

    Article  CAS  PubMed  Google Scholar 

  18. Dezfulian M, Hatheway CL, Yolken RH, Bartlett JG (1984) Enzyme-linked immunosorbent assay for detection of Clostridium botulinum type A and type B toxins in stool samples of infants with botulism. J Clin Microbiol 20:379–383

    CAS  PubMed Central  PubMed  Google Scholar 

  19. Doellgast GJ, Triscott MX, Beard GA, Bottoms JD (1994) Enzyme-linked immunosorbent assay-enzyme-linked coagulation assay for detection of antibodies to Clostridium botulinum neurotoxins A, B, and E and solution-phase complexes. J Clin Microbiol 32:851–853

    CAS  PubMed Central  PubMed  Google Scholar 

  20. Doellgast GJ, Brown JE, Koufman JA, Hatheway CL (1997) Sensitive assay for measurement of antibodies to Clostridium botulinum neurotoxins A, B, and E: use of hapten-labeled-antibody elution to isolate specific complexes. J Clin Microbiol 35:578–583

    CAS  PubMed Central  PubMed  Google Scholar 

  21. Fach P, Gibert M, Griffais R, Guillou JP, Popoff MR (1995) PCR and gene probe identification of botulinum neurotoxin A-, B-, E-, F-, and G-producing Clostridium spp. and evaluation in food samples. Appl Environ Microbiol 61:389–392

    CAS  PubMed Central  PubMed  Google Scholar 

  22. McGrath S, Dooley JS, Haylock RW (2000) Quantification of Clostridium botulinum toxin gene expression by competitive reverse transcription-PCR. Appl Environ Microbiol 66:1423–1428

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  23. Berry PR, Rodhouse JC, Hughes S, Bartholomew BA, Gilbert RJ (1998) Evaluation of ELISA, RPLA, and Vero cell assays for detecting Clostridium perfringens enterotoxin in faecal specimens. J Clin Pathol 41:458–461

    Article  Google Scholar 

  24. Szilagyi M, Rivera VR, Neal D, Merrill GA, Poli MA (2000) Development of sensitive colorimetric capture ELISA for Clostridium botulinum neurotoxin serotypes A and B. Toxicon 38:381–389

    Article  CAS  PubMed  Google Scholar 

  25. Szabo EA, Pemberton JM, Desmarchelier PM (1993) Detection of the genes encoding botulinum neurotoxin types A to E by the polymerase chain reaction. Appl Environ Microbiol 59:3011–3020

    CAS  PubMed Central  PubMed  Google Scholar 

  26. Ashton AC, Crowther JS, Dolly JO (1985) A sensitive and useful radioimmunoassay for neurotoxin and its haemagglutinin complex from Clostridium botulinum. Toxicon 2:235–246

    Article  Google Scholar 

  27. Wong R (2002) The effect of adulterants on urine screen for drugs of abuse: detection by an on-site dipstick device. Am Clin Lab 21:37–39

    CAS  PubMed  Google Scholar 

  28. Tsay A, Williams L, Mitchell EB, Chapman MD (2002) A rapid test for detection of mite allergens in homes. Clin Exp Allergy 32:1596–1601

    Article  CAS  PubMed  Google Scholar 

  29. Shyu RH, Tang SS, Chiao DJ, Hung YW (2010) Gold nanoparticle-based lateral flow assay for detection of staphylococcal enterotoxin B. Food Chem 118:462–466

    Article  Google Scholar 

  30. Peng DP, Hu SS, Hua Y, Xiao YC, Li ZL, Wang XL (2007) Comparison of a new gold-immunochromatographic assay for the detection of antibodies against avian influenza virus with hemagglutination inhibition and agar gel immunodiffusion assays. Vet Immunol Immunopathol 117:17–25

    Article  CAS  PubMed  Google Scholar 

  31. Al-Yousif Y, Anderson J, Chard-Bergstrom C, Kapil S (2002) Development, evaluation, and application of lateral-flow immunoassay (immunochromatography) for detection of rotavirusin bovine fecal samples. Clin Diagn Lab Immunol 9:723–724

    PubMed Central  PubMed  Google Scholar 

  32. Qian S, Bau HH (2003) A mathematical model of lateral flow bioreactions applied to sandwich assays. Anal Biochem 322:89–98

    Article  CAS  PubMed  Google Scholar 

  33. Chiao DJ, Wey JJ, Shyu RH, Tang SS (2008) Monoclonal antibody-based lateral flow assay for detection of botulinum neurotoxin type A. Hybridoma 27:31–35

    Article  CAS  PubMed  Google Scholar 

  34. Woody MA, DasGupta BR (1988) Fast protein liquid chromatography of botulinum neurotoxin types A, B and E. J Chromatogr 430:279–289

    Article  CAS  PubMed  Google Scholar 

  35. Gram HC (1884) “Über die isolierte Färbung der Schizomyceten in Schnitt-und Trockenpräparaten” (in German). Fortschritte der Medizin 2:185–189 (English translation in: Brock TD (1999) Milestones in microbiology, 2 ed. ASM Press, pp 215–218)

    Google Scholar 

  36. Doellgast GJ, Triscott MX, Beard GA, Bottoms JD, Cheng T, Roh BH, Roman MG, Hall PA, Brown JE (1993) Sensitive enzymelinked immunosorbent assay for detection of Clostridium botulinum neurotoxins A, B, and E using signal amplification via enzyme-linked coagulation assay. J Clin Microbiol 31:2402–2409

    CAS  PubMed Central  PubMed  Google Scholar 

  37. Ferreira JL, Maslanka S, Johnson E, Goodnough M (2003) Detection of botulinal neurotoxins A, B, E, and F by amplified enzyme-linked immunosorbent assay: collaborative study. J AOAC Int 86:314–331

    CAS  PubMed  Google Scholar 

  38. Boroff DA, Chu-Chen G (1973) Radioimmunoassay for type A toxin of Clostridium botulinum. Appl Microbiol 25:545–549

    CAS  PubMed Central  PubMed  Google Scholar 

  39. Shyu RH, Shyu HF, Liu HW, Tang SS (2002) Colloidal gold-based immunochromatographic assay for detection of ricin. Toxicon 40:255–258

    Article  CAS  PubMed  Google Scholar 

  40. Diederen BMW, Peeters MF (2006) Evaluation of rapid U Legionella Plus Test, a new immunochromatographic assay for detection of Legionella pneumophila serogroup 1 antigen in urine. Eur J Clin Microbiol Infect Dis 25:733–735

    Article  CAS  PubMed  Google Scholar 

  41. Ferreira JL, Eliasberg SJ, Harrison MA, Edmonds P (2001) Detection of preformed type A botulinal toxin in hash brown potatoes by using the mouse bioasssay and a modified ELISA test. J AOAC Int 84:1460–1464

    CAS  PubMed  Google Scholar 

  42. Rivera VR, Gamez FJ, Keener WK, White JA, Poli MA (2006) 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. Anal Biochem 353:248–256

    Article  CAS  PubMed  Google Scholar 

  43. Shone C, Wilton-Smith P, Appleton N, Hambleton P, Modi SG, Melling J (1985) Monoclonal antibody-based immunoassay for type A Clostridium botulinum toxin is comparable to the mouse assay. Appl Environ Microbiol 50:63–67

    CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by a Grant (102-B6) from the Institute of Preventive Medicine, National Defense Medical Center.

Conflict of interest

There are no financial or other relations that could lead to a conflict of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rong-Hwa Shyu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yeh, CT., Liang, CC., Yang, CM. et al. Comparison of immunostrips with mouse bioassay and bacterial culture in detecting botulinum toxins in bottles from suspected Taiwan high-speed rail bomber. Forensic Toxicol 32, 258–265 (2014). https://doi.org/10.1007/s11419-014-0235-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11419-014-0235-8

Keywords

Navigation