C CFU assay

Yeast Infection Free Forever

Effective Home Remedies for Candida Albicans

Get Instant Access

1 Add 0.1 ml of Candida suspension (containing 1 x 10" yeasts) to wells containing washed monocytes/macrophages (effector-to-target ratio 10:l)a and to six wells not containing macrophages to be used as control cultures.

2 Add 0.1 ml of complete medium to each well to yield a final volume of 0.2 ml/well.

3 Incubate the plates containing macrophages and Candida for 3 h at 37°C.

4 Remove the plates from the incubator and shake vigorously on a shaking platform.

5 Stop the intracellular Candida killing by lysing the phagocytic cells with 20 |xl/well of a solution of 10% Triton X-100 in water.

6 Carefully mix each well by pipetting and make serial dilutions in distilled water (from 1:20 to 1:100 final dilution).

7 Plate 0.1 ml of serial dilutions on Sabouraud dextrose agar (triplicate samples).

8 Count the surviving colony-forming units (CFU) after 24 h of incubation at 37°Cand compare them to control cultures consisting of C. albicans incubated without effector cells.

9 Express the results as the percentage of anti-Candida activity according to the formula:

{1 - (CFU of experimental group/CFU of control culture)} x 100.

3 Different effector-to-target ratios should be tested (e.g. from 20:1 to 2.5:1). Usually, the number of target cells is kept constant and an effector-to-target ratio titration Is achieved by varying the number of effector cells.

To optimize this test, it is important to use an agerminative strain of C. albicans that grows as a pure yeast form in vitro at 28°C or 37°C in conventional media. This will avoid bias due to hyphae formation during the incubation (49).

4.2.3 Extracellular killing of C. Albicans

Protocol 9 describes a method for assessment of extracellular killing of Candida (hyphal form) using the (3-(4,5-dimethy 1thiazol-2-yl)-2,5-di-phenyIterrazolium) MTT colorimetric assay.

Was this article helpful?

0 0
How To Cure Yeast Infection

How To Cure Yeast Infection

Now if this is what you want, you’ve made a great decision to get and read this book. “How To Cure Yeast Infection” is a practical book that will open your eyes to the facts about yeast infection and educate you on how you can calmly test (diagnose) and treat yeast infection at home.

Get My Free Ebook

Post a comment