Efficient DNA Fingerprinting of Clostridium botulinum Types A, B, E, and F by Amplified Fragment Length Polymorphism Analysis
- 1 March 2005
- journal article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 71 (3) , 1148-1154
- https://doi.org/10.1128/aem.71.3.1148-1154.2005
Abstract
Amplified fragment length polymorphism (AFLP) analysis was applied to characterize 33 group I and 37 group II Clostridium botulinum strains. Four restriction enzyme and 30 primer combinations were screened to tailor the AFLP technique for optimal characterization of C. botulinum . The enzyme combination HindIII and HpyCH4IV, with primers having one selective nucleotide apiece (Hind-C and Hpy-A), was selected. AFLP clearly differentiated between C. botulinum groups I and II; group-specific clusters showed C. botulinum strains. In addition, group-specific fragments were detected in both groups. All strains studied were typeable by AFLP, and a total of 42 AFLP types were identified. Extensive diversity was observed among strains of C. botulinum type E, whereas group I had lower genetic biodiversity. These results indicate that AFLP is a fast, highly discriminating, and reproducible DNA fingerprinting method with excellent typeability, which, in addition to its suitability for typing at strain level, can be used for C. botulinum group identification.Keywords
This publication has 54 references indexed in Scilit:
- An Improved Amplified Fragment Length Polymorphism (AFLP) Protocol for Discrimination of Listeria IsolatesSystematic and Applied Microbiology, 2003
- Characterisation of Persistent and Sporadic Listeria monocytogenes Strains by Pulsed-Field Gel Electrophoresis (PFGE) and Amplified Fragment Length Polymorphism (AFLP)Systematic and Applied Microbiology, 2003
- Lesson of the week: Wound botulism associated with subcutaneous drug useBMJ, 2002
- Comparison of Pulsed-Field Gel Electrophoresis and Amplified Fragment Length Polymorphism Techniques for Investigating Outbreaks of Enteritis Due to CampylobactersJournal of Clinical Microbiology, 2002
- Molecular Fingerprinting of Clostridium difficile Isolates: Pulsed-Field Gel Electrophoresis versus Amplified Fragment Length PolymorphismJournal of Clinical Microbiology, 2002
- Multiplex PCR Assay for Detection and Identification ofClostridium botulinumTypes A, B, E, and F in Food and Fecal MaterialApplied and Environmental Microbiology, 2001
- Differentiation of Campylobacter species by AFLP fingerprintingMicrobiology, 2001
- Evaluation of the DNA fingerprinting method AFLP as a new tool in bacterial taxonomyMicrobiology, 1996
- AFLP: a new technique for DNA fingerprintingNucleic Acids Research, 1995
- Genetic interrelationships of saccharolytic Clostridium botulinum types B, E and F and related clostridia as revealed by small-subunit rRNA gene sequencesFEMS Microbiology Letters, 1993