AIM: To isolate and identify pathogenic fungi in patient’s specimens as an aid in the diagnosis of fungal infections (mycoses)
MATERIAL/APPARATUS: Patient’s specimen, incubator, inoculating loop, Bunsen burner, S+C tube, S+C+A tube, lactophenol cotton blue reagent, glass slide, cover slip, microscope.
S = Sabouraud’s dextrose agar (SDA)
C = Chloramphenicol
A = Actidine (cyclohexamide)
- Label the S+C and S+C+A tubes with the patient’s biodata (e.g. name, date of culturing, laboratory number, and type of specimen).
- Collect the specimen using a sterilized inoculating loop.
- Inoculate the specimen on the slope and butt of the S+C and S+C+A tubes.
- Incubate the tubes in the incubator at 28oC for about 21 days.
- Check the incubated tubes for fungal growth at different time intervals before the 21 days elapses.
- After the 21 days, bring out the S+C and S+C+A tubes from the incubator.
- Make wet preparations of the fungal growths in both tubes on a clean glass slide using the lactophenol cotton blue stain.
- Cover the prepared slide with a cover slip and view with the ×10 and ×40 objective lens of the microscope.
REPORTING OF THE RESULT:
The name of the fungus responsible for the disease condition of the patient is reported after a thorough microscopical investigation is carried out. Familiarity with fungal elements and structures also helps in this process.