TABLE 5.
Comparison of tetraplex real-time PCRs developed previously by Ngobeni et al. and in this study
| Feature | Result for tetraplex real-time PCR developed by: |
|
|---|---|---|
| Ngobeni et al. (19) | Ali and Roy (this study) | |
| Simultaneous detection of E. histolytica, E. dispar, E. moshkovskii, and E. bangladeshi | Yes | Yes |
| Target DNA | 18S SSU rDNA | 18S SSU rDNA |
| Real-time PCR platform | Hydrolysis probes (TaqMan) | Hydrolysis probes (TaqMan) |
| Primers | One pair, common to all four Entamoeba species | Four pairs, each specific to an Entamoeba species |
| Probes | Four, each specific to an Entamoeba species | Four, each specific to an Entamoeba species |
| Sensitivity | Unknown | Well defined; capable of detecting ameba DNA originating from a single trophozoite |
| Amplicon size | Larger (250 bp) | Smaller (132–145 bp) |
| Use in conventional PCR format? | No | Yes |
| Detection limit for disproportionately infected mixed infections | Not tested | Can detect E. histolytica in the presence of up to 10-fold more DNA from one or more Entamoeba species |
| Compared with another established detection method? | No | Yes, compared with a diagnostic duplex real-time PCR for E. histolytica and E. dispar that is routinely used at U.S. CDC |