Species | Purification/Concentration | Nucleic Acid Extraction Kit | References |
Entamoeba histolytica | 1) Filtration using 0.3 mm sieve to remove coarse materials. 2) Filtration of the liquid through 0.22 µm nitrate cellulose membrane filters. 3) Centrifugation of the filtered specimens at 4500 G for 30 minutes. | DNA isolation kit (Macherey Nagel GmBH, Germany) | [45] |
Entamoeba histolytica | 1) Filtration using 800 µm filter paper to remove sediments. 2) Centrifuged at 5000 G for 10 minutes. | QIAamp DNA mini kit (QIAGEN, Hilden, Germany) | [64] |
Entamoeba dispar | 1) Centrifuge at 4000 G at 4˚C for 30 minutes. 2) Resuspension in phosphate buffer solution (pH 7.4) and additional centrifugation at 4300 G at 4˚C for 15 minutes. 3) Resuspension in 280 µL of Buffer AL (Qiagen, Hilden, Germany) and 20 µL proteinase-K (Qiagen) and incubate at 56˚C for 60 minutes. 4) Repeat at −80˚C for 30 minutes. | MagNa Pure LC Total Nucleic Acid Isolation Kit with the MagNa Pure LC 2.0 Instrument (Roche Diagnostics, Basel, Switzerland) | [65] |
E. histolytica & E. dispar | 1) Filtration of 5 L through 1.2 µm filters. 2) Wash concentrated samples with 50 mL phosphate buffer solution. 3) Centrifuge at 1500 G for 5 minutes. 4) Perform immunomagnetic separation and sucrose flotation on the produced supernatant. | QIAamp DNA minikit | [22] |