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J Clin Microbiol. Sep 1993; 31(9): 2255–2262.
PMCID: PMC265744

Gene probes for identification of the botulinal neurotoxin gene and specific identification of neurotoxin types B, E, and F.


A polymerase chain reaction method was developed for the specific detection of the botulinum neurotoxin (BoNT) gene of Clostridium botulinum. Degenerate oligonucleotide primers, designed from the nucleotide sequence of the heavy chain of the BoNT gene, amplified a specific fragment of approximately 1.1 kb from strains of C. botulinum toxin types A, B, E, F, and G and neurotoxin-producing strains of Clostridium barati and Clostridium butyricum, but no fragment was obtained from nontoxigenic strains. The fragments amplified from several strains of C. botulinum types B, E, and F were cloned in Escherichia coli and their nucleotide sequences were determined. Sequences within this region were used to design oligonucleotide probes specific for BoNT type B (BoNT/B), BoNT/E, and BoNT/F genes. An additional probe was designed for the detection of the BoNT/F gene of C. barati, which differed in sequence from BoNT/F genes of both proteolytic and nonproteolytic strains of C. botulinum.

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Selected References

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  • Aureli P, Fenicia L, Pasolini B, Gianfranceschi M, McCroskey LM, Hatheway CL. Two cases of type E infant botulism caused by neurotoxigenic Clostridium butyricum in Italy. J Infect Dis. 1986 Aug;154(2):207–211. [PubMed]
  • Binz T, Kurazono H, Popoff MR, Eklund MW, Sakaguchi G, Kozaki S, Krieglstein K, Henschen A, Gill DM, Niemann H. Nucleotide sequence of the gene encoding Clostridium botulinum neurotoxin type D. Nucleic Acids Res. 1990 Sep 25;18(18):5556–5556. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Boroff DA, Chu-Chen G. Radioimmunoassay for type A toxin of Clostridium botulinum. Appl Microbiol. 1973 Apr;25(4):545–549. [PMC free article] [PubMed]
  • Campbell KD, East AK, Thompson DE, Collins MD. Studies on the large subunit ribosomal RNA genes and intergenic spacer regions of non-proteolytic Clostridium botulinum types B, E and F. Res Microbiol. 1993 Mar-Apr;144(3):171–180. [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • East AK, Richardson PT, Allaway D, Collins MD, Roberts TA, Thompson DE. Sequence of the gene encoding type F neurotoxin of Clostridium botulinum. FEMS Microbiol Lett. 1992 Sep 15;75(2-3):225–230. [PubMed]
  • Hall JD, McCroskey LM, Pincomb BJ, Hatheway CL. Isolation of an organism resembling Clostridium barati which produces type F botulinal toxin from an infant with botulism. J Clin Microbiol. 1985 Apr;21(4):654–655. [PMC free article] [PubMed]
  • Hauser D, Eklund MW, Kurazono H, Binz T, Niemann H, Gill DM, Boquet P, Popoff MR. Nucleotide sequence of Clostridium botulinum C1 neurotoxin. Nucleic Acids Res. 1990 Aug 25;18(16):4924–4924. [PMC free article] [PubMed]
  • Hauser D, Gibert M, Boquet P, Popoff MR. Plasmid localization of a type E botulinal neurotoxin gene homologue in toxigenic Clostridium butyricum strains, and absence of this gene in non-toxigenic C. butyricum strains. FEMS Microbiol Lett. 1992 Dec 1;78(2-3):251–255. [PubMed]
  • Hutson RA, Thompson DE, Collins MD. Genetic interrelationships of saccharolytic Clostridium botulinum types B, E and F and related clostridia as revealed by small-subunit rRNA gene sequences. FEMS Microbiol Lett. 1993 Mar 15;108(1):103–110. [PubMed]
  • Lewis GE, Jr, Kulinski SS, Reichard DW, Metzger JF. Detection of Clostridium botulinum type G toxin by enzyme-linked immunosorbent assay. Appl Environ Microbiol. 1981 Dec;42(6):1018–1022. [PMC free article] [PubMed]
  • McCroskey LM, Hatheway CL, Fenicia L, Pasolini B, Aureli P. Characterization of an organism that produces type E botulinal toxin but which resembles Clostridium butyricum from the feces of an infant with type E botulism. J Clin Microbiol. 1986 Jan;23(1):201–202. [PMC free article] [PubMed]
  • McCroskey LM, Hatheway CL, Woodruff BA, Greenberg JA, Jurgenson P. Type F botulism due to neurotoxigenic Clostridium baratii from an unknown source in an adult. J Clin Microbiol. 1991 Nov;29(11):2618–2620. [PMC free article] [PubMed]
  • Miller CA, Anderson AW. Rapid detection and quantitative estimation of type A botulinum toxin by electroimmunodiffusion. Infect Immun. 1971 Aug;4(2):126–129. [PMC free article] [PubMed]
  • Poulet S, Hauser D, Quanz M, Niemann H, Popoff MR. Sequences of the botulinal neurotoxin E derived from Clostridium botulinum type E (strain Beluga) and Clostridium butyricum (strains ATCC 43181 and ATCC 43755). Biochem Biophys Res Commun. 1992 Feb 28;183(1):107–113. [PubMed]
  • Szabo EA, Pemberton JM, Desmarchelier PM. Specific detection of Clostridium botulinum type B by using the polymerase chain reaction. Appl Environ Microbiol. 1992 Jan;58(1):418–420. [PMC free article] [PubMed]
  • Thompson DE, Brehm JK, Oultram JD, Swinfield TJ, Shone CC, Atkinson T, Melling J, Minton NP. The complete amino acid sequence of the Clostridium botulinum type A neurotoxin, deduced by nucleotide sequence analysis of the encoding gene. Eur J Biochem. 1990 Apr 20;189(1):73–81. [PubMed]
  • Whelan SM, Elmore MJ, Bodsworth NJ, Atkinson T, Minton NP. The complete amino acid sequence of the Clostridium botulinum type-E neurotoxin, derived by nucleotide-sequence analysis of the encoding gene. Eur J Biochem. 1992 Mar 1;204(2):657–667. [PubMed]
  • Whelan SM, Elmore MJ, Bodsworth NJ, Brehm JK, Atkinson T, Minton NP. Molecular cloning of the Clostridium botulinum structural gene encoding the type B neurotoxin and determination of its entire nucleotide sequence. Appl Environ Microbiol. 1992 Aug;58(8):2345–2354. [PMC free article] [PubMed]

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