TABLE 4.

Genetic typing tools applied for Clostridium botulinum groups I and II

Method	Discrimination of groups I and II (level)	Effort and time required to completion from pure colony	Reproducibility	Special feature	Application(s)	Reference(s)
PFGE	Strain	Laborious, 5-7 days	Excellent	DNA degradation by clostridial endonucleases may disturb analysis	Epidemiological studies, comparison of clinical isolates	96, 110, 130, 131, 160, 161
rRNA gene sequencing	Group	Moderate, 3-4 days	Excellent		Taxonomy	33, 41, 107, 108
Ribotyping	Group	Laborious, 3-5 days (for     automated ribotyping,     easy, 1 day)	Moderate	Can be automated (RiboPrinter)	Phylogeny, species/group identification	97, 191
AFLP	Strain and group	Easy, 3 days	Excellent		Epidemiological studies, comparison of clinical isolates, phylogeny	120
RAPD	Strain	Easy, 1-2 days	Low	Can be performed with crude cell lysate (although this further reduces reproducibility)	Rapid comparison of clinical isolates, not suited to interlaboratory comparison of strains	109
Rep-PCR	Group I, toxinotype; group II, strain	Easy, 2 days	Moderate	Not well suited to gram-positive bacteria	Rapid comparison of clinical isolates	109