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]