Table 1

Comparison of Common Bacterial Typing Techniques by Relative Discriminatory Power, Reproducibility, Repeatability, and Whether They Give Information on Dispersed or Focal Parts of the Genome, Time Required and Cost

Typing Technique
Relative discriminatory power
Relative repeatability
Relative reproducibility
Dispersed or focal parts of the genome*
Days required post culture
Relative Cost**
Notes

Sequencing of entire genome
High
High
High
Entire genome
Months to years
Very high

Comparative hybridization against array containing entire gene sequence
High
Medium to high
Medium to high
Dispersed
Weeks to months
High
Microarrays are increasingly available for human pathogens – not all genes will be present in the sequenced strain
Direct sequencing of one or more genetic regions
Moderate to high (depends on gene choice)
High
High
Focal if only one region
2–3
Equipment: Medium to High
Labor & Supplies: Medium to High
Initial selection of target genes might be time consuming.
Multilocus sequence typing (MLST)
Moderate to high (depends on gene choice)
High
High
Dispersed
3+
Equipment: Medium to High
Labor & Supplies: High
Initial selection of target genes might be time consuming. Species specific.
Binary typing (presence/absence of selected genes or alleles across the genome)
Moderate to high (depends on gene choice)
High
Potentially High
Dispersed (if chose different genes across the genome)
2–3
Equipment: medium
Labor & Supplies: Medium
Reliability dependent on DNA yield and purity
Pulsed-field gel electrophoresis (PFGE)
Moderate to high (depends on number of bands observed)
Medium=> High (depending on species)
Medium =>High
Dispersed
3
Equipment: High
Labor & Supplies: High
Discrimination depends on type and number of enzymes selected.
Restriction fragment length polymorphism (RFLP)
Moderate to High (depends on number of bands observed)
Medium=>High
Medium
Dispersed
1–3
Medium

Amplification of a single target gene specific to a pathogen
Moderate to high (depends on gene choice)
High
Medium=>High
Focal
<1
Equipment: Low to Medium
Labor & Supplies: Low

Amplified fragment length polymorphism (AFLP)
Moderate to high
High
Medium=>High
Dispersed
2
Equipment: Low to Medium
Labor & Supplies: Low

Automated ribotyping
Moderate
High
High
Focal
1
Equipment: High
Labor & Supplies: High
Works for most bacterial species
Ribosomal RNA gel electrophoresis
Moderate
High
High
Focal
1
Equipment: Low
Labor & Supplies: Medium

Targeting known repetitive gene sequences (enterobacterial repetitive intergenic consensus sequences (ERIC), repetitive extragenic palindromic sequences (REP), DRE (double repetitive element), BOX, insertional sequence (IS), polymorphic GC-rich repetitive sequences (PGRS))
Low to moderate
Medium
Low
Generally dispersed
1
Equipment: Low to Medium
Labor & Supplies: Low
Patterns vary with equipment used
Random primers (randomly amplified polymorphic DNA (RAPD), arbitrary primed PCR (AP-PCR))
Low to moderate
Low
Low
Dispersed
1
Equipment: Low to Medium
Labor & Supplies: Low
Patterns vary with equipment used
Restriction endonuclease on a single amplified product
Low to moderate (depends on amplicon)
High
High
Focal
1–2
Equipment: Low to Medium
Labor & Supplies: Low

Plasmid profiles
Low
High
Medium
Focal
1
Equipment: Low
Labor & Supplies: Low


*Focal corresponds to interrogating a single loci. Dispersed means multiple loci are interrogated.

**Per isolate costs in US dollars in 2005, assuming all equipment are available, and the investigator has access to automatic sequencing, for PCR reactions are ~$5, PFGE~$20, MLST ~$140, comparative hybridization~$1000 to $2000 and total genomic sequencing (assuming a strain has already been sequenced)~$100,000 to $500,000.

Note: For a summary and details of these techniques, and assessments of repeatability and reproducibility, see Tenover, 1997 [1], Gurtler and Mayall 2001 [2] and VanBelkum, 2003 [3]. In general, sequence-based methods are most repeatable and reproducible. Gel-based methods are less so, because of the inherent variability of the technique.

Foxman et al. Epidemiologic Perspectives & Innovations 2005 2:10   doi:10.1186/1742-5573-2-10