Investigation of dermatological specimens for superficial mycoses

Microbiology

Dermatophytes are the most common cause for superficial mycoses in hair, nail and skin.

Dermatophytes can be divided into three groups: anthropophilic, zoophilic, and geophilic. Anthropophilic dermatophytes are passed from human to human and are the most common in the community. Zoophilic or animal acquired infections are usually sporadic. Infections with geophilic dermatophytes are most often acquired following a close association with soil or from an animal itself infected by soil contact. Infection is diagnosed by observing the presence of fungal hyphae in skin, hair or nail specimens. However, it is important to culture the material to determine the infecting genus and species. This is done to ensure selection of the most appropriate therapy and in order to trace its likely epidemiology.

Dermatophyte (otherwise known as ringworm) infections are usually referred to as tinea followed by the Latin name of the body area involved. The most common dermatophyte infections are tinea pedis in adults (athlete’s foot) which may also include tinea unguium (nail infection), and tinea capitis (scalp ringworm) in children.

Infection by dermatophytes is cutaneous and generally restricted to the non-living cornified layers in patients who are immunocompetent. This is because the dermatophyte group of fungi are generally unable to penetrate tissues which are not fully keratinised (i.e. deeper tissues and organs). However, reactions to such infections can range from mild to severe depending upon the host’s immune response, the virulence of the infecting species, the site of infection and environmental factors.

The dermatophyte group of fungi are classified in three genera: Epidermophyton species, Microsporum species and Trichophyton species.

Non-dermatophytes;

There are few non-dermatophyte moulds that can infect otherwise healthy skin and these include Scytalidium dimidiatum, Scytalidium hyalinum (a white variant of S. dimidiatum), Phaeoannellomyces werneckii and Piedraia hortae. Non-dermatophyte moulds, including those above, can infect nails damaged by physical trauma, disease or pre-existing infection with a dermatophyte. There are many non-dermatophyte moulds that have been implicated in nail infection, therefore isolation of a mould from a nail specimen should only be reported if certain strict criteria are met because contamination of nail samples with mould spores is common. A non-dermatophyte mould accounts for the diagnosis in less than 5% of infected nails.

Sample Type:

See above precautions.

Temperature: Ambient Room Temperature

Turnaround Time:

Microscopy results available after 3 days.

Culture results available following 3 weeks incubation.

*Excludes Saturdays, Sundays and Bank Holidays

Sample Stability:

Specimens should be kept at ambient room temperature and transported and processed as soon as possible although, provided the samples are kept dry, the fungus will remain viable for several months.

Instrument / Procedure:

Manual Microscopy & Culture

Units:
Reference Range:
Precautions:

PLEASE NOTE: Samples and accompanying relevant patient/ isolate data maybe referred for confirmatory or further laboratory testing to Reference laboratories. Relevant Public Health departments may also be notified IF a notifiable disease is identified under the Infectious Diseases (Amendment) Regulations 2020 (S.I. No. 53/2020).

Preparation of patient: Collect specimens before antimicrobial therapy where possible.

Precautions: Specimens should be collected into folded paper squares secured and placed in a plastic bag or in commercially available packets designed specifically for the collection and transport of skin, nail and hair samples.

Skin;

Patients’ skin and nails can be swabbed with 70% alcohol prior to collection of the specimen, this is especially important if creams, lotions or powders have been applied. The edges of skin lesions yield the greatest quantities of viable fungus. Lesions should be scraped with a blunt scalpel blade. If insufficient material can be obtained by scraping, then sticky tape can be pressed on the lesion and transferred to a clean glass slide for transport to the laboratory (’stripping’).

Nails;

Good nail samples are difficult to obtain. It should be specified whether the sample is from the fingernails or toenails. Material should be taken from any discoloured, dystrophic or brittle parts of the nail. The affected nail should be cut as far back as possible through the entire thickness and should include any crumbly material. Nail drills, scalpels and nail elevators may be helpful but must be sterilized between patients. When there is superficial involvement (as in white superficial onychomycosis) nail scrapings may be taken with a curette. If associated skin lesions are present samples from these are likely to be infected with the same organism and are more likely to give a positive culture.

Hair;

Samples from the scalp should include skin scales and plucked hairs or hair stumps. Cut hairs are not suitable for direct examination as the infected area is usually close to the scalp surface. Plastic hairbrushes, scalp massage pads or plastic toothbrushes may be used to sample scalps for culture where there is little obvious scaling, but such samples do not replace a scraping for direct examination.

Specimens should be kept at room temperature and transported and processed as soon as possible although, provided the samples are kept dry, the fungus will remain viable for several months.

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SAMPLE REQUIREMENTS FOR COAGULATION TESTS

PROCEDURE

Sample Requirements and Collection

  • Patients should be relaxed pre-venepuncture. Excessive stress and exercise will increase FVIII, vWF antigen and fibrinolysis. Venous occlusion should be avoided.
  • Difficult venepuncture with trauma may lead to platelet activation with release of PF4 fromalpha granules.
  • Venous blood should be collected into coagulation tubes containing Sodium Citrate 3.2%, 0.105M, 3ml.
  • Specimens must be mixed immediately post venepuncture to avoid clot activation, by GENTLY inverting the tubes 5 to 10 times.
  • The ratio of whole blood to anticoagulant is crucial. Under-filled specimens will not be processed as over- or under-filled tubes can adversely affect results.
  • Any warfarin treatment should be mentioned on the request form.

Transportation and Storage

  • PT/INR specimens should ideally be analysed within 12 hours of collection and transported to the laboratory at room temperature.
  • APTT and Fibrinogen should ideally be analysed within 4 hours of collection. Where this is not possible please centrifuge at room temperature (RT) @3000rpm (1500g) for at least 15 minutes, and then carefully remove the plasma from the cells, transfer to a fresh plastic plain tube and freeze at -20oC.
  • Non-frozen coagulation specimens should be transported at RT ASAP to avoid deterioration of labile factors V and VIII.
  • Collection of blood through intravenous lines that have been previously flushed with heparin should be avoided. In the event blood is drawn from an indwelling catheter, the line should be flushed with 5ml of saline, and the first 5ml of blood or 6 times the line volume be drawn off and discarded before coagulation tube is filled.

Plasma Sample Stability (CLSI H21-A5)

  • PT 24 hours @ RT or 2 weeks @ -20oC
  • APTT 4 hours @ RT or 2 weeks @ -20oC & 12 Months @ -70oC
  • Fibrinogen - 4 hours @ RT `

ESR Ref Ranges
Units of Measurement
MALE
FEMALE
>50 Years
mm/hr
0 - ≤12
0 - ≤15
<50 Years
mm/hr
0 - ≤8
0 - ≤10
Analyte
Units of Measurement
MALE
FEMALE
WBC
10^9/L
4.0 - 10.0
4.0 - 10.0
RBC
10^12/L
4.5 - 5.5
3.8 - 4.8
HB
g/dL
13.0 - 17.0
12.0 - 15.0
HCT
L/L
0.400 - 0.500
0.360 - 0.460
MCV
fL
83 - 101
83 - 101
MCH
pg
27 - 32
27 - 32
MCHC
g/dL
31.5 - 34.5
31.5 - 34.5
PLT
10^9/L
150 - 410
150 - 410
MPV
fL
N/A
N/A
RDW
%
11.6 - 14.0
11.6 - 14.0
#Neut
10^9/L
2.0 - 7.0
2.0 - 7.0
#Lymph
10^9/L
1.0 - 3.0
1.0 - 3.0
#Mono
10^9/L
0.2 - 1.0
0.2 - 1.0
#Eos
10^9/L
0.02 - 0.50
0.02 - 0.50
#Baso
10^9/L
0.02 - 0.10
0.02 - 0.10
Analyte
Units of Measurement
MALE
FEMALE
WBC
10^9/L
Up to 1 year (6.00-16.00)
Up to 6 years (5.00-15.00)
Up to 12 years (5.00-13.00)
Up to 18 years (3.88-10.49)
Up to 1 year (6.00-16.00)
Up to 6 years (5.00-15.00)
Up to 12 years (5.00-13.00)
Up to 18 years (3.88-10.49)
RBC
10^12/L
Up to 1 year (3.90-5.10)
Up to 12 years (4.00-5.20)
Up to 18 years (4.28-5.59)
Up to 1 year (3.90-5.10)
Up to 12 years (4.00-5.20)
Up to 18 years (3.73-5.02)
HB
g/dL
Up to 1 year (11.1-14.1)
Up to 6 years (11.0-14.0)
Up to 12 years (11.5-15.5)
Up to 18 years (13.5-17.2)
Up to 1 year (11.1-14.1)
Up to 6 years (11.0-14.0)
Up to 12 years (11.5-15.5)
Up to 18 years (11.3-15.2)
HCT
L/L
Up to 1 year (0.300-0.380)
Up to 6 years (0.340-0.400)
Up to 12 years (0.350-0.450)
Up to 18 years (0.381-0.499)
Up to 1 year (0.300-0.380)
Up to 6 years (0.340-0.400)
Up to 12 years (0.350-0.450)
Up to 18 years (0.323-0.462)
MCV
fL
Up to 1 year (72.0-84.0)
Up to 6 years (75.0-87.0)
Up to 12 years (77.0-95.0)
Up to 18 years (83.1-99.1)
Up to 1 year (72.0-84.0)
Up to 6 years (75.0-87.0)
Up to 12 years (77.0-95.0)
Up to 18 years (83.1-99.1)
MCH
pg
Up to 1 year (25.0-29.0)
Up to 6 years (24.0-30.0)
Up to 12 years (25.0-33.0)
Up to 18 years (28.3-33.9)
Up to 1 year (25.0-29.0)
Up to 6 years (24.0-30.0)
Up to 12 years (25.0-33.0)
Up to 18 years (28.3-33.9)
MCHC
g/dL
Up to 1 year (32.0-36.0)
Up to 6 years (31.0-37.0)
Up to 12 years (31.0-37.0)
Up to 18 years (32.1-36.6)
Up to 1 year (32.0-36.0)
Up to 6 years (31.0-37.0)
Up to 12 years (31.0-37.0)
Up to 18 years (32.1-36.6)
PLT
10^9/L
Up to 1 year (200-550)
Up to 6 years (200-490)
Up to 12 years (170-450)
Up to 18 years (164-382)
Up to 1 year (200-550)
Up to 6 years (200-490)
Up to 12 years (170-450)
Up to 18 years (164-382)
RDW
%
No separate paediatric ranges
No separate paediatric ranges
#Neut
10^9/L
Up to 1 year (1.00-7.000)
Up to 6 years (1.50-8.00)
Up to 12 years (2.00-8.00)
Up to 18 years (1.56-6.52)
Up to 1 year (1.00-7.000)
Up to 6 years (1.50-8.00)
Up to 12 years (2.00-8.00)
Up to 18 years (1.56-6.52)
#Lymph
10^9/L
Up to 1 year (3.50-11.00)
Up to 6 years (6.00-9.00)
Up to 12 years (1.00-5.00)
Up to 18 years (1.01-3.13)
Up to 1 year (3.50-11.00)
Up to 6 years (6.00-9.00)
Up to 12 years (1.00-5.00)
Up to 18 years (1.01-3.13)
#Mono
10^9/L
Up to 1 year (0.20-1.00)
Up to 6 years (0.20-1.00)
Up to 12 years (0.20-1.00)
Up to 18 years (1.01-3.13)
Up to 1 year (0.20-1.00)
Up to 6 years (0.20-1.00)
Up to 12 years (0.20-1.00)
Up to 18 years (1.01-3.13)
#Eos
10^9/L
Up to 12 years (0.10-1.00)
Up to 18 years (0.05-0.51)
Up to 12 years (0.10-1.00)
Up to 18 years (0.05-0.51)
#Baso
10^9/L
Up to 1 day (0.00-0.64)
Up to 7 days (0.00-0.25)
Up to 14 years (0.00-0.23)
Up to 18 years (0.02-0.15)
Up to 1 day (0.00-0.64)
Up to 7 days (0.00-0.25)
Up to 14 years (0.00-0.23)
Up to 18 years (0.02-0.15)
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