|
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 |
|||||||