Detection of Clostridium botulinum type F using the polymerase chain reaction

Ferreira, Joseph; Hamdy, M. K.; McCay, Steven G.; Hemphill, Mark L.; Kirma, Nameer B.; Baumstark, Barbara R.

doi:10.1006/mcpr.1994.1053

Cited by 19 publications

(6 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The amino acid sequences corresponding to these gene sequences are further referred to as BTxF1 (Langeland), BTxF2 (202F), BTxF3 (CDC 3281), and BTxF4 (C. barati ATCC 43756), respectively. For the C. botulinum type F strain investigated here, strain Langeland, the neurotoxin gene sequence was partly confirmed before, in two independent studies [45,46]. 10 In addition, the gene sequence of the NTNH of strain Langeland is known (NCBInr X99064; [47]).…”

Section: Available Sequence Informationmentioning

confidence: 54%

Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

Baar¹,

Hulst²,

Jong³

et al. 2004

Journal of Chromatography A

View full text Add to dashboard Cite

In a follow-up of the earlier characterisation of botulinum toxins type A and B (BTxA and BTxB) by mass spectrometry (MS), types C, D, E, and F (BTxC, BTxD, BTxE, BTxF) were now investigated. Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. Biologically active BTxC, BTxD, BTxE, and BTxF are comprised of a protein complex of the respective neurotoxins with non-toxic non-haemagglutinin (NTNH) and, sometimes, specific haemagglutinins (HA). These protein complexes were observed in mass spectrometric identification. The BTxC complex, from Clostridium botulinum strain 003-9, consisted of a 'type C1 and D mosaic' toxin similar to that of type C strain 6813, a non-toxic non-hemagglutinating and a 33 kDa hemagglutinating (HA-33) component similar to those of strain C-Stockholm, and an exoenzyme C3 of which the sequence was in full agreement with the known genetic sequence of strain 003-9. The BTxD complex, from C. botulinum strain CB-16, consisted of a neurotoxin with the observed sequence identical with that of type D strain BVD/-3 and of an NTNH with the observed sequence identical with that of type C strain C-Yoichi. Remarkably, the observed protein sequence of CB-16 NTNH differed by one amino acid from the known gene sequence: L859 instead of F859. The BTxE complex, from a C. botulinum isolated from herring sprats, consisted of the neurotoxin with an observed sequence identical with that from strain NCTC 11219 and an NTNH similar to that from type E strain Mashike (1 amino acid difference with observed sequence). BTxF, from C. botulinum strain Langeland (NCTC 10281), consisted of the neurotoxin and an NTNH; observed sequences from both proteins were in agreement with the gene sequence known from strain Langeland. As with BTxA and BTxB, matrix-assisted laser desorption/ionisation (MALDI) MS provided provisional identification from trypsin digest peptide maps and liquid chromatography-electrospray (tandem) mass spectrometry (LC-ES MS) afforded unequivocal identification from amino acid sequence information of digest peptides obtained in trypsin digestion.

show abstract

Section: Available Sequence Informationmentioning

confidence: 54%

Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

Baar¹,

Hulst²,

Jong³

et al. 2004

Journal of Chromatography A

View full text Add to dashboard Cite

show abstract

“…PCR-based assays have been reported for BoNT gene fragments using conventional block based assays with gel electrophoresis for detection of amplified products (Campbell et al, 1993;Szabo et al, 1993;Ferreira et al, 1994;Franciosa et al, 1994Franciosa et al, , 1998Fach et al, 1995;Takeshi et al, 1996;Aranda et al, 1997;Hyytia et al, 1998;Alsallami and Kotlowski, 2001;Dahlenborg et al, 2001;Lindström et al, 2001;Nevas et al, 2002;Craven et al, 2002). These assays either detected a single toxin type or suffered from a lack of specificity due to low melting temperatures of the primers, some requiring subsequent use of hybridisation probes to confirm the specific toxin type.…”

Section: Discussionmentioning

confidence: 99%

Development and Application of Real-Time PCR Assays to Detect Fragments of theClostridium botulinumTypes A, B, and E Neurotoxin Genes for Investigation of Human Foodborne and Infant Botulism

Akbulut¹,

Grant²,

McLauchlin³

2004

Foodborne Pathogens and Disease

View full text Add to dashboard Cite

Real-time PCR assays for detection of Clostridium botulinum neurotoxin (BoNT) gene fragments specific to BoNTA, B, and E were developed as alternatives to the mouse bioassay. The expected specificities of the PCR assays were demonstrated by in silico analysis as well as empirical testing of target DNA extracted from 83 pure cultures of C. botulinum, and 44 bacteria from other species. The sensitivities of the assays were found to be equivalent to 16, 10, and 141 genomes for BoNT A, B, and E, respectively. The assays were shown to be applicable to both purified DNA, as well as crude DNA extracted from cultures and enrichment broths. The assays were evaluated using DNA extracted directly from clinical and food specimens as well as from inoculated broths using material collected from seven confirmed and one suspected case of botulism. The appropriate BoNT genes were detected in material from seven of the eight cases of botulism and provided a supportive diagnosis faster than the conventional bioassay. These assays have already proven useful for pubic health microbiological investigation of suspected cases of human botulism by substantially improving the diagnostic process.

show abstract

“…botulinum genomic DNA was isolated as described previously [11]. botulinum genomic DNA was isolated as described previously [11].…”

Section: Ampli¢cation Of Overlapping Toxin Type B Gene Fragmentsmentioning

confidence: 99%

“…C. botulinum genomic DNA was isolated as described previously [11]. The Roche Molecular Biochemicals (Indianapolis, IN, USA) Expand kit was used to amplify overlapping type B toxin gene fragments ( Fig.…”

Section: Ampli¢cation Of Overlapping Toxin Type B Gene Fragmentsmentioning

confidence: 99%

Characterization of six type A strains ofClostridium botulinumthat contain type B toxin gene sequences

Kirma

Ferreira

Baumstark

2004

FEMS Microbiology Letters

Self Cite

View full text Add to dashboard Cite

Six Clostridium botulinum isolates exhibiting type A toxicity as measured by the mouse bioassay were found to contain both type A and type B neurotoxin DNA sequences. The six strains were divided into three groups based on the DNA sequence of the type B neurotoxin gene. Members of each group exhibited 100% sequence identity over the 3876 bp type B toxin open reading frame. The type B toxin sequence of all groups differed at more than 60 positions when compared to the BGB control strain.

show abstract

Detection of Clostridium botulinum type F using the polymerase chain reaction

Cited by 19 publications

References 0 publications

Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

Development and Application of Real-Time PCR Assays to Detect Fragments of theClostridium botulinumTypes A, B, and E Neurotoxin Genes for Investigation of Human Foodborne and Infant Botulism

Characterization of six type A strains ofClostridium botulinumthat contain type B toxin gene sequences

Contact Info

Product

Resources

About