Sop Od Dermatophytosis
Sop Od Dermatophytosis
1. Purpose- This procedures describe the process for isolation and identification of
species of dermatophytes from the samples taken from infected skin , hair and nails.
2.Scope- This SOP provides guidlines for sample collection, storage and transportation along
with the procedures for identification of fungal species.
1.Sample Collection
A) Skin Scrapings:
C) Hair:
Infected hair will be removed by plucking with epilating forceps
or
scraping the scalp with a blunt scalpel
IDENTIFICATION OF DERMATOPHYTES
Identification of Dermatophytes
Microscopy Culture
Colour Texture
Surface Reverse
1. DIRECT MICROSCOPY:
Transfer small quantity of the specimen with a loop or the tip of a scalpel
into the KOH drop
Then examine under the microscope 10x lens, then 40x objective lens
(condensor, iris and diaphragm should be sufficiently closed)
Observation
1. Examine the clear specimen under low power (10X or 20X objective).
Scan the entire cover slip from end to end in a zigzag fashion.
2. If any fungal elements are suspected, examine under high power (40X
objective).
3. Reduce the light coming into the condenser while examining at high
power.
4. Look for branching hyphae, type of branching, the colour, septation
and thickness of hyphae, and spores.
Name Signature
Prepared by Dr. Ananya Verma
Checked by Dr. Suneel Kumar Ahirwar
Approved by Dr. Anita Mutha
The plates are incubated aerobically at 25-30ºc for upto 21 days and
are checked daily for appearance of fungal colonies.
Name Signature
Prepared by Dr. Ananya Verma
Checked by Dr. Suneel Kumar Ahirwar
Approved by Dr. Anita Mutha
PRINCIPLES: A tease prepration is the quickest and most common technique for
mounting fungi for microscopic examination.Since the mould’s growth is teased apart
with dissecting needles, conidia or spores may be disloged from the conidiogenous or
sporogenous cells.
Precise identification is done by examine the colony under microscope.
MATERIALS:
1.Long handled inoculating needle
2.two dissecting needles
3.Lactophenol
4.Clean microscopic slide and cover slip
5.Clear fingernail polish
PROCEDURE:
Remove aseptically a small portion of growth midway between the colony center and
edge. Place the removed colony on a drop of lactophenol cotton blue on a slide
Tease the fungus using a pair of dissecting needles so as to have a thin spread out
Gently place a cover slip at the edge of the drop of mounting fluid.
Avoid trapping air bubbles. Excess of lactophenol can by wiped out using a blotting
paper
For preserving the mount, seal the edges of coverslip with nail polish/varnish
Examine the slide under the microscope using 10x objective lens, then 40x objective
lens.
Name Signature
Prepared by Dr. Ananya Verma
Checked by Dr. Suneel Kumar Ahirwar
Approved by Dr. Anita Mutha
STANDARD OPERATING PROCEDURE
SOP NO. : 5 Date of issue:
Procedure for isolation Effective date :
Department of and identification of Page 8 of 10
Microbiology dermatophytes
4. SLIDE CULTURE:
Whenever it is difficult to identify moulds with tease mounts, slide cultures can be put
up. Nutritionally deficient media like corn meal agar is good for enhancing sporulation.
PROCEDURE:
Aseptically cut 1 cm square agar blocks from CMA
Transfer very small amount of colony to the four sides of the agar block
With sterile forceps, place the coverslip on the inoculated agar block
Add 1 – 1.5 mL, of sterile water to the petri dish; humid atmosphere does not allow the
agar block to dry out
(5-20% glycerine can be added to the sterile water to prevent condensation of moisture
on the slide)
Place the slide culture in a petridish canister and incubate in the dark.
Slide culture is ready to be taken down when mature conidia or spores are observed.
Result:
After performing the above procedures ,most common fungal species from different
infected sites like skin , hair and nails are identified.
Name Signature
Prepared by Dr. Ananya Verma
Checked by Dr. Suneel Kumar Ahirwar
Approved by Dr. Anita Mutha