Veterinary Dermatology 2005, 16, 102107

Development of an enzyme-linked immunosorbant assay (ELISA)

for the serodiagnosis of canine dermatophytosis caused by
Microsporum canis

Blackwell Publishing, Ltd.

ANDREA PEANO, LUISA RAMBOZZI and MARIA G. GALLO

Dipartimento di Produzioni Animali, Epidemiologia ed Ecologia, Facolt di Medicina Veterinaria, Universit
degli Studi di Torino, Via Leonardo da Vinci, 44, Grugliasco (Turin), Italy
(Received 24 August 2004; accepted 1 February 2005)

Abstract In dogs, dermatophytosis should be considered in any case of alopecic, papular or pustular lesion. The
aim of this study was to develop an enzyme-linked immunosorbant assay (ELISA) as an aid in the diagnosis of
canine dermatophytosis. The antigen used was a whole fungal extract obtained from an isolate of Microsporum
canis cultured on a liquid medium from the parasitized hair of a cat with patches of alopecia. To assess the ELISA
performances, sera from 18 dogs with dermatophytosis caused by M. canis (group A, n = 18), 20 dogs with skin
diseases other than dermatophytosis and 22 healthy dogs (group B, n = 42) were tested. Four further animals were
tested: three with dermatophytosis caused by M. gypseum and one by T. mentagrophytes. A significant difference
(P < 0.01, Wilcoxons test, w = 364) was found between IgG-specific levels of sera of recently M. canis-infected
dogs (infection < 15 days) and controls (although three dogs had negative titres at this stage). A highly significant
difference (P < 0.001, w = 462) was noted between controls and dogs with infection of longer duration (> 30
days). All dogs had positive titres at this stage. A highly significant correlation (P < 0.001, Spearmans test,
rho = 0.86) between duration of infection and IgG concentration was noted. The test has good sensitivity (83.3%)
and high specificity (95.2%) but some dogs retained positive titres after elimination of infection. The sensitivity
is higher than that of direct microscopic hair examination and similar to that of fungal culture with DTM
(dermatophyte test medium).

I NTRO D U CTI ON
Canine dermatophytosis is a skin disease caused by
keratinophilic fungi belonging to the genera Microsporum
and Trichophyton. Microsporum canis, M. gypseum and
T. mentagrophytes cause the great majority of infections in clinical cases. Various reports have demonstrated
that M. canis is a common cause of canine dermatophytosis. There is great variation in the proportion in
which these three fungi occur in different parts of the
world, and their incidence and prevalence varies with
climate and natural reservoirs.1 Dermatophytes are
transmitted by contact with infected hair and scale or
fungal elements on animals, in the environment, or on
fomites.2 They are highly contagious and readily transmissible to humans. In dogs, the most consistent clinical sign is one or more circular patches of alopecia.
However, signs and symptoms are highly variable and
depend on hostfungus interaction and, thus, on the
degree of inflammation.2 The differential diagnosis for
dermatophyosis is extensive and this disease should be
considered in any case of alopecic, papular or pustular
lesion.3 The diagnosis can be challenging and fungal
tests are very useful. Woods lamp examination for
fluorescence causes only certain strains of M. canis to
produce a positive yellow-green colour on infected
Correspondence: Luisa Rambozzi, Dipartimento di Produzioni
Animali, Epidemiologia ed Ecologia, Via Leonardo da Vinci 44,
10095 Grugliasco, Turin, Italy. E-mail: luisa.rambozzi@unito.it
102

hair. Only about 50% of M. canis infections fluoresce.

Several major pitfalls exist in the use and interpretation
of Woods lamp results.2 The sensitivity of direct microscopic hair examination varies from 37%4 to 70%2 in dogs
and cats; its specificity, evaluated in humans, is about
60%.5 The fungal culture of affected hair and scales is
the most reliable diagnostic method but requires
experience and time. A diagnostic kit based upon colour
change of the culture medium (DTM, dermatophyte
test medium) has been used in recent years. False positive and false negative results are possible and sensitivity
corresponds to 82%.6 Biopsy findings may be a definitive proof of true infection but histopathological aspects
are as variable as the clinical lesions and are less sensitive than culture.2 Despite the superficial nature of the
infection, dermatophytoses evoke cellular and humoral
immune responses.7 The production of specific antibodies
has been demonstrated in humans, cats, guinea pigs,
rabbits and dogs.812 At the moment no commercial
test is available for the serological diagnosis of dermatophytosis. The aim of this study was to develop an
enzyme-linked immunosorbant assay (ELISA) for the
serological diagnosis of dermatophytosis in dogs.

M AT E R IA L S A N D M E T H O D S
Preparation of the antigen
An isolate of M. canis was cultured on selective
medium agar plates (Mycobios Selective Agar, Biolife
2005 European Society of Veterinary Dermatology

Serodiagnosis of canine dermatophytosis

s.r.l., Milan, Italy) from the parasitized hair of a cat
with patches of alopecia. The fungus was identified by
the morphology of the thallus and the microscopic
appearance of macro- and microconidia,13 then subcultured on Sabourauds agar and finally in Sabouraud
broth (Sabouraud Broth, Biolife s.r.l.) for 15 days
at 25 C. The mycelium was harvested by filtration
through filter paper, washed in distilled water, frozen
and, after thawing, re-suspended in phosphate-buffered
saline solution (PBS), and homogenized and sonicated
with three 1-min cycles at 4 C, to destroy the hyphas.
The suspension was centrifuged at 2000 g for 30 min
and the supernatant taken and re-centrifuged at
20 000 g for 60 min at 4 C. Total protein concentration in the fungal extract was determined by the use of
a protein assay (DC Protein Assay, BIO-RAD, Hercules, CA, USA). The product was divided into aliquots
of 150 L and stored at 20 C until use.

Serum samples
Positive and negative controls were represented by
two pools: five subjects with dermatophytosis due to
M. canis identified by fungal culture and five subjects
with no historical or clinical evidence of dermatophytosis. The serum was separated by centrifugation and
stored at 20 C until assayed. Using a pool reduces
the possibility of technical errors and ensures uniformity in the different phases of the study by creating
reference data for repetitively tested samples. These
sera were also used for the evaluation of the test procedure by intra- and inter-assay precision.
To assess the test performances, blood samples were
collected from 60 subjects: 18 dogs with clinical evidence
of active dermatophytosis caused by M. canis, confirmed
by fungal culture (group A, n = 18); 20 dogs with skin
diseases other than dermatophytosis and 22 dogs with
no cutaneous signs or historical evidence of dermatophytosis (group B, n = 42). In dogs with skin disease,
dermatophytosis was ruled out by negative fungal
cultures from hair and crusts.
These subjects comprised nine dogs with pyoderma,
four with flea allergy, two with sarcoptic mange, one
with demodicosis, one with vasculitis, one with contact
hypersensitivity, and two with food hypersensitivity.
For dogs of group A, the correlation between the
duration of the infection, obtained by anamnesis, and
IgG concentration was evaluated.
In addition, four animals with skin lesions caused by
dermatophytes other than M. canis (three M. gypseum
and one T. mentagrophytes) were tested.

ELISA assays
Assays were performed in polystyrene microtitre plates
(Sigma-Aldrich Corporation, Saint Louis, MO, USA)
coated, for 1 h at 37 C and then overnight at 4 C,
with 100 L per well of 15 g ml1 crude antigen solution in PBS. The last row was left free of antigen as
control. After washing with PBS mixed with Tween 20,
0.05%, pH 7.2 (PBST), the plates were saturated with
5% nonfat dried milk in PBST for 1 h at 37 C. Triplicate

103

serum samples were diluted to 1 : 200 in PBST and

added for 1 h at 37 C to the antigen-coated wells. After
washing, 100 L of 1 : 10 000-diluted antidog IgG peroxidase conjugate (Anti-Dog IgG-peroxidase conjugate,
Sigma-Aldrich Corporation) was added to each well.
After a further 1-h incubation at 37 C the plates were
washed and a peroxidase substrate (Sigma FAST ophenylenediamine dihydrolchoride tablet sets, SigmaAldrich Corporation) was added. The plates were kept
in the dark for 15 min and the resultant optical density
(OD) of the well contents read at 450 nm using an
automated plate reader (Plate Reader, DAS s.r.l.,
Palombara Sabina, Rome, Italy).

Analysis of results
Optical density was defined as the difference between
the adsorbance mean for each triplicated serum tested
and the control wells. Results were then expressed as
OD percentages (OD%) obtained as follows: (OD
sample OD negative control)/(OD positive control OD
negative control). Reproducibility of the test procedure
was determined by intra-assay (CV1) and inter-assay
(CV2) coefficients of variance (CV = standard deviation/
mean) by measuring five replicates each of positive and
negative controls in a single plate and subsequently in
five separate assays. Following the method of Visual
Inspection of Frequency Distributions14,15 the cut-off
value was fixed at the intersection point of the OD%
value distribution curves of the two groups of animals
tested. Comparing these ELISA results against the
gold standard represented by the fungal culture of hair
and scales, the subjects were divided into four categories: true positives (TP) (culture positive, ELISA positive),
true negatives (TN) (culture negative, ELISA negative), false negatives (FN) (culture positive, ELISA
negative) and false positives (FP) (culture negative,
ELISA positive). ELISA sensitivity (Se) and specificity
(Sp) were calculated as follows:
Se = (TP/TP + FN) 100; Sp = (TN/TN + FP) 100.

Statistical evaluation
The results were analysed using Wilcoxon and Spearman correlation tests with R (1-4-1 version) and
EpiInfo 6 software.

R E S U LT S
The absolute OD values of the negative and positive
control sera in a typical experiment were approximately 0.46 (range 0.430.47) and 1.29 (range 1.23
1.3), respectively. The CV1 was 4.1 and 7.6% for the
positive and negative controls, respectively, while the
CV2 was 5.6 and 7.5%. In group A, the mean of the
OD% values corresponded to 112.6 (SD: 52.7); in
group B to 26 (SD: 26). The cut-off value was fixed at
OD% 70 (Fig. 1), giving the test a sensitivity of 83.3%
and a specificity of 95.2% (Table 1). A highly significant
difference (P < 0.001, Wilcoxons test, w = 682) existed

2005 European Society of Veterinary Dermatology, Veterinary Dermatology, 16, 102107

104

A Peano et al.

Figure 1. Distribution of the ELISA optical

density percentage values of sera from dogs
affected by M. canis (group A) (grey bars)
and from control dogs (group B) (white
bars).

Positive serology
Negative serology
Total

Table 1. ELISA results: cases and controls.

OD% values > 70 were considered positive

Group A*

Group A

Group B

Positive culture

Positive culture

Negative culture

Total

5
3
8

10
0
10

2
40
42

17
43
60

*Infection < 15 days, infection > 30 days.

between the group A and group B OD% values. For

dogs of group B with skin diseases other than dermatophytosis the mean of the OD% values was 23.7, while
for dogs of group B without skin lesions it was to 28. No
significant difference (P = 0.45, Wilcoxons test, w = 190)
existed between these two subgroups.
The OD% mean for M. canis-infected dogs with recent
lesions (less than 15 days, n = 8) was 66.6 (SD: 46.9);
that of subjects with longer infection (more than 30 days,
n = 10) was 149.3 (SD: 12.4). The test applied to the
former group had a sensitivity of 62.5%; for the latter
it corresponded to 100% (Table 1).
For the four animals with dermatophytosis caused
by dermatophytes other than M. canis, the OD% values
were as follows: 157 (T. mentagrophytes), 99, 15 and
124 (M. gypseum).
The seronegative dermatophytic dogs (three M. canis
and one M. gypseum) had OD% values above the cutoff point when re-tested after variable periods, all of them
still having lesions positive to cultural examination. Five
dogs, having recovered from infection (no lesions and with
negative culture) were also re-tested after variable periods.
Three of them (two M. canis and one M. gypseum) had
OD% below the cut-off point; the others (one M. canis
and one M. gypseum) still had OD% above the cut-off.

D ISCU SSIO N
This study confirmed the production of a humoral
response in canine dermatophytosis, as already

demonstrated in cats. 16 The test discriminated well

between dermatophytic animals and controls, with
good sensitivity (83.3%) and high specificity (95.2%).
The test has a higher sensitivity than that of direct
microscopic hair examination carried out in dogs, cats
and humans2,4,5,17 and similar to that of fungal culture
with DTM.6 Moreover, it requires only a short time with
no particular ability or experience. On DTM, some
dermatophyte isolates do not produce the initial red
colour change, which is highly dependent on incubation temperature and number of infected hairs deposited on the reactive substrate.2,18 Three dogs of group A
had no measurable antibodies in spite of a positive
culture (Table 1). To rule out laboratory errors as the
cause of false negative results, the same negative sera of
the three dogs were re-tested and found to have the
same results. As the three animals were not puppies, the
cause for the discordant results could not be attributed
to an immature immune system. Another potential
cause for the falsely negative titres prior treatment
with corticosteroids was excluded by anamnesis. When
re-tested after variable periods, the three dermatophytic
seronegative dogs had high measurable antibody levels.
Specific IgG production, even if occurring later, clearly
differentiated all three infected dogs from negative controls. With regard to dogs for which the initial ELISA
result was positive, it is not clear if IgG were produced
during the incubation phase or after lesions had
appeared. During experimental infection by M. canis
in guinea pigs, IgG levels were detectable 14 days after
the lesions appeared,10,19 while in cats IgG were high

2005 European Society of Veterinary Dermatology, Veterinary Dermatology, 16, 102107

Serodiagnosis of canine dermatophytosis

when lesions first appeared in some animals and
4 weeks later in others.20 In dogs, specific IgG were
detected 1421 days after an experimental M. canis
infection12 while the incubation period before the
appearance of lesions has been shown to be 1012
days.21 In our study, OD% values were highly variable
in the early stages of infection and a significant correlation (P < 0.001, Spearmans test, rho = 0.86) between
duration of infection, obtained by anamnesis, and IgG
concentration was found. In another study, IgG
concentration was higher in dogs naturally infected
1753 days earlier than in dogs experimentally infected
1421 days earlier.12 The difference between IgG
values of newborn T. mentagrophytes-infected rabbits and
controls became significant (P < 0.01) when the subjects
were 7 weeks old and highly significant (P < 0.001) at
11 weeks.11 Our study showed that a significant difference (P < 0.01, Wilcoxons test, w = 364) existed between
the OD% values of recently M. canis-infected dogs
(infection < 15 days) and controls, while a highly significant difference (P < 0.001, w = 462) was noted
between controls and animals with infection of longer
duration (> 30 days). For these latter dogs, ELISA
sensitivity was equivalent to 100%. An ELISA test has
presented the same sensitivity value in chronic dermatophytic cats.9 In the rabbits described above,11 ELISA
sensitivity was 96% 11 weeks after infection. A window
probably exists between onset of clinical signs and
seroconversion during dermatophytosis in some dogs.
Sampling of the falsely negative dogs seemed therefore
to have occurred during this window. Variations in IgG
increase may reflect the complex interaction of fungal
antigens with the immune system, whose reactivity is
partially responsible for various cutaneous signs. However, in our study, humoral response intensity and
precocity did not seem to correlate with severity of the
clinical signs or the extent of the lesions, confirming
what has already been reported for dogs affected by
dermatophytosis.12 Likewise, in asymptomatic dermatophytic cats, OD% values were the same as those for
subjects with full-blown infection,9 whereas in chronically T. mentagrophytes-infected humans a correlation
between the extent of the lesions and specific IgG and
IgA has been reported.22 In dogs with a longer infection and with serious clinical signs, high IgG titres may
reflect persistent antigen stimulation by the fungus.
Nevertheless, it is possible that the high and persistent
IgG concentrations represent an ineffective and aberrant immune response, which does not allow complete
elimination of the fungus.23 Chronically infected humans
often maintain high specific IgG titres, failing to
develop a cellular protective immune response. IgG
titres generally decrease in patients who recover from
infection.8 It remains to be proven whether antibody
titres persist as a result of the presence of antigens produced by the fungus, or whether this presence is the
result of the continuing antibody titres.
All the animals were tested using a single cat-derived
M. canis strain as antigen. This seems to confirm
the serological cross-reactivity of different strains of

105

M. canis and may even suggest cross-reactivity with the

other two major dermatophytes of pets, in agreement
with previous studies concerning dermatophytosis
in cats.9,16 In fact, of the four dogs with skin lesions
caused by dermatophytes other than M. canis, three
(two M. gypseum and one T. mentagrophytes) were
ELISA positive and the remaining one (M. gypseum)
became positive later. Our study did not include sufficient patients infected with M. gypseum and T. mentagrophytes to enable definite claims to be made. Further
study to establish the diagnostic reliability for the latter
two fungal species would allow a rapid diagnosis of all
cases, irrespective of the dermatophyte involved in the
skin pathology. However, it should be noted that even
if therapy does not require identification of the species,2 this becomes important for a correct approach to
all aspects regarding the treatment of a dermatophytic
dog (identification of the source of infection, environmental contamination, transmission to other animals
and humans, etc.).
In our study ELISA specificity was high even though
variable antidermatophyte IgG levels were found in
many controls. In humans and cats this has been
thought to depend on previous dermatophyte
infections or ubiquitous environmental fungi crossreactivity.8,16 In healthy cats and humans a correlation
between age and antidermatophyte IgG levels was
found. This probably reflected the increase of exposure
to environmental fungal antigens during their lifetime.7,16 In young cats kept isolated as negative controls
in vaccination trials, slight increases in antidermatophyte IgG and IgM concentrations were in any case
noted.24 Some human antigens, such as intercellular
glycoproteins and an erythrocyte antigen, have been
shown to cross-react with dermatophyte antigens.8 In
dogs, serological cross-reactivity with Candida albicans
and Malassezia pachydermatis was not found, while the
role of Aspergillus fumigatus has still not been satisfactorily determined.12 Our study did not find a correlation
between age and OD% values (P = 0.051, Spearmans
test, rho = 0.3) in the control group. This seems to
indicate that in most controls these IgG levels were due
to a misdiagnosed dermatophytosis not reported in
anamnesis. This is in agreement with analogous situations found in epidemiological surveys in humans.8 For
this reason we tested ELISA specificity by monitoring
IgG levels in five dogs after they had recovered from
dermatophytosis. Three of them had OD% values below
the chosen cut-off (two M. canis and one M. gypseum),
one (M. canis) had a decreased value but above the cutoff, while the last (M. gypseum) had an increased OD%
value. All these animals had completely recovered and
had a negative fungal culture. No correlation was
found with the length of time from recovery, as the
three ELISA negative animals had recovered 3
4 months before and the one with increased OD%
levels 5 months before, whereas the final dog had
recovered 1 year before. In cats, 8 months after recovering from a dermatophyte infection, some animals
had stable ELISA values, while the values of the others

2005 European Society of Veterinary Dermatology, Veterinary Dermatology, 16, 102107

106

A Peano et al.

had halved.9,24,25 Moreover, no correlation was found

between a humoral response and recovery from
infection in cats, whereas a correlation between the
development of a cellular immune response and the
elimination of fungi was reported in cats, guinea pigs
and humans.8,10,20 In a recent study, Garcia26 reported
the potential use of an immunodiagnostic method for
monitoring two dogs affected by dermatophytosis due
to M. canis. Our results show that the decrease in IgGspecific levels seems variable and not correlated to aetiological recovery. As serology could detect either active
or prior dermatophyte infection and as only relatively
few recovered animals were available for testing, work
is continuing to ascertain how long antibodies persist
after the elimination of dermatophytes from affected
animals. Information on this aspect will be necessary
to assess the full potential of a serological test for the
diagnosis and follow-up of canine dermatophytosis.

ACKN OWLEDGE ME NT
The authors thank Dr Anna Rita Molinar for valuable
technical help.

R E FEREN CES
1. Foil CS. Dermatophytosis. In: Greene CE ed. Infectious
Diseases of the Dog and Cat. Philadelphia: W.B.
Saunders, 1998: 362 70.
2. Miller WH, Scott DW, Griffin CE. Small Animal Dermatology, 6th edn. Philadelphia: W.B. Saunders, 2000.
3. Noli C, Scarampella F. Dermatologia del Cane e del
Gatto. Milano: Poletto ed, 2002.
4. Peano A, Gallo MG, Peyrot R. Epidemiologia, clinica
delle dermatofitosi negli animali domestici e risultati di
unindagine svolta su cani e gatti della citt di Torino
negli anni 1996 1999. Proceedings of the 5th National
Meeting of the Federazione Italiana di Micopatologia
Umana e Animale, 2000: 16.
5. Miller MA, Hodgson Y. Sensitivity and specificity of
potassium hydroxide smears of skin scraping for the
diagnosis of Tinea pedis. Archives of Dermatology 1993;
129: 510 11.
6. Carroll HF. Evaluation of dermatophyte test medium for
diagnosis of dermatophytosis. Journal of the American
Veterinary Medical Association 1974; 165: 192 5.
7. Woodfolk JA, Slunt JB, Deuell B et al. Definition of a
Trichophyton protein associated with delayed hypersensitivity in humans. Evidence for immediate (IgE and IgG4)
and delayed hypersensitivity to a single protein. Journal
of Immunology 1996; 156: 1695 701.
8. Woodfolk JA, Platts-Mills TA. The immune response to
dermatophytes. Research in Immunology 1998; 149:
436 45.
9. Mignon BR, Coignoul F, Leclipteux T et al. Histopathological pattern and humoral immune response to a crude
exo-antigen and purified keratinase of Microsporum canis
in symptomatic and asymptomatic infected cats. Medical
Mycology 1999; 37: 1 9.

10. Mignon BR, Leclipteux T, Focant C et al. Humoral and

cellular immune response to a crude exo-antigen and
purified keratinase of Microsporum canis in experimentally infected guinea pigs. Medical Mycology 1999; 37:
1239.
11. Zrimsek P, Kos J, Pinter L et al. Detection by ELISA of
the humoral immune response in rabbits naturally
infected with Trichophyton mentagrophytes. Veterinary
Microbiology 1999; 70: 778.
12. Pinter L, Noble WC, Ellis J et al. The value of enzymelinked immunosorbant assay (ELISA) in the serodiagnosis of canine dermatophytosis due to Microsporum
canis. Veterinary Dermatology 1992; 3: 6570.
13. Rebell G, Taplin D. Dermatophytes: Their Recognition
and Identification. Coral Gables, Florida: University of
Miami Press, 1974.
14. International Organisation for Standardisation (ISO).
Quality management and quality assurance standards.
Part III. Guidelines for the Application of ISO 9001:1994
to the Development, Supply, Installation and Maintenance of Computer Software. ISO 9003. Geneva: ISO,
1997: 132.
15. Jacobson RH. Validation of serological assays for
diagnosis of infectious diseases. Revue Scientifique et
Technique Office Internationale des Epizooties 1998;
17: 46986.
16. Sparkes AH, Stokes CR, Gruffyd-Jones TJ. Humoral
immune responses in cats with dermatophytosis. American
Journal of Veterinary Research 1993; 54: 186973.
17. Aho R, Oksanen A, Pihlaja O. Direct microscopic examination of hair samples. Suomen Elainlaakarilehti 1993;
99: 1658.
18. Guillot J, Latie L, Deville M et al. Evaluation of dermatophyte test medium RapidVet-D. Veterinary Dermatology
2001; 12: 1237.
19. Pier AC, Hodges AB, Lauze JM et al. Experimental
immunity to Microsporum canis and cross reactions with
other dermatophytes of veterinary importance. Journal
of Medical and Veterinary Mycology 1995; 33: 937.
20. Sparkes AH, Stokes CR, Gruffyd-Jones TJ. Experimental
Microsporum canis infection in cats: correlation between
immunological and clinical observations. Journal of
Medical and Veterinary Mycology 1995; 33: 17784.
21. Khosla R, Rai P, Gupta P. Experimental Microsporum
canis infection in dogs. Indian Journal of Animal
Sciences 1989; 59: 2215.
22. Honbo S, Jones HE, Artis WM. Chronic dermatophyte
infection: evaluation of the Ig class-specific antibody
response reactive with polysaccharide and peptide antigens derived from Tricophyton mentagrophytes. Journal
of Investigative Dermatology 1984; 82: 28790.
23. DeBoer DJ, Moriello KA. Humoral and cellular
immune responses to Microsporum canis in naturally
occurring feline dermatophytosis. Journal of Medical
and Veterinary Mycology 1993; 31: 12132.
24. DeBoer DJ, Moriello KA. The immune response to
Microsporum canis induced by a fungal cell wall vaccine.
Veterinary Dermatology 1994; 5: 4755.
25. Sparkes AH, Gruffyd-Jones TJ, Stokes CR. Acquired
immunity in experimental feline Microsporum canis
infection. Research in Veterinary Science 1996; 61: 1658.
26. Garcia ME, Guedeja-Marron J, Lazaro M et al.
Monitoring of canine dermatophytosis by an immunodiagnostic method. Veterinary Record 2004; 154: 116 17.

2005 European Society of Veterinary Dermatology, Veterinary Dermatology, 16, 102107

Serodiagnosis of canine dermatophytosis

107

Rsum Chez le chien, une dermatophytose doit tre suspecte dans tous les cas de lsions alopciques, papuleuses et pustuleuses. Le but de cette tude tait de dvelopper un test ELISA pour le diagnostic de dermatophytose canine. Lantigne utilis tait un extrait de champignon obtenu dun isolat de culture de M. canis provenant
dun chat parasit qui souffrait dune alopcie. Pour dterminer lefficacit du test ELISA, le srum de 18 chiens
dermatophytose M. canis (groupe A, n = 18), de 20 chiens dermatose autre et de 22 chiens sains (group B,
n = 42) ont t tests. Quatre autres chiens ont t tudis (trois prsentant une dermatophytose M. gypseum
et un T. mentagrophytes. Une diffrence significative (P < 0.01, Wilcoxon test, w = 364) a t note pour les taux
dIgG spcifiques des chiens rcemment infests par M. canis (infection < 15 jours) et ceux du groupe contrle
(bien que trois chiens prsentent des titres ngatifs ce stade), et une diffrence hautement significative (P < 0.001,
w = 462) a t note entre les animaux du groupe contrle et ceux prsentant une infestation chronique (> 30
jours). Tous les chiens avaient des titres positifs cette date. Une corrlation hautement significative (P < 0.001,
Spearman test, rho = 0.86) entre la dure de linfection et les concentrations en IgG a t observe. Le test avait
une bonne sensibilit (83.3%) et une spcificit leve (95.2%) mais certains chiens ont prsent des titres positifs
aprs gurison de linfection. La sensibilit est meilleure que celle de lexamen direct des poils et semblable la
culture fongique avec un DTM (Dermatophyte Test Medium).
Resumen En perros con lesiones alopcicas, papulares o pustulares se debe considerar siempre la dermatofitosis. El objetivo de este estudio fue desarrollar un anlisis de ELISA como ayuda en el diagnstico de la dermatofitosis canina. El antgeno utilizado fue un extracto fngico entero obtenido de un cultivo de M. canis en
un medio lquido de pelo infectado de un gato con focos alopcicos. Para evaluar el funcionamiento del ELISA,
se probaron sueros de 18 perros con dermatofitosis causada por M. canis (grupo A, n = 18), 20 perros con
enfermedades cutneas con excepcin de dermatofitosis y 22 perros sanos (grupo B, n = 42). Cuatro animales
ms fueron probados, tres con dermatofitosis causada por M. gypseum y uno por T. mentagrophytes. Exista una
diferencia significativa (P < 0.01, test de Wilcoxon, w = 364) entre los niveles de IgG-especficos de sueros de perros recientemente infectados con M. canis (infeccin < 15 das) y los controles (aunque tres perros tenan ttulos
negativos en esta fase), mientras que se observ una diferencia altamente significativa (P < 0.001, w = 462) entre
los controles y los perros con infeccin de una duracin ms larga (> 30 das). Todos los perros tenan ttulos
positivos en esta fase. Se observ una correlacin altamente significativa (P < 0.001, test de Spearman,
rho = 0.86) entre la duracin de la infeccin y la concentracin de IgG. La prueba tiene la buena sensibilidad
(83.3%) y alta especificidad (95.2%) pero algunos perros conservan ttulos positivos despus de la eliminacin
de la infeccin. La sensibilidad es ms alta que la del examen microscpico directo del pelo y similar a la del cultivo
fngico con DTM (Dermatophyte Test Medium).

Zusammenfassung Bei Hunden sollte bei jedem einzelnen Fall einer alopezischen, papulren und pustulsen
Lsion eine Dermatophytose in Betracht gezogen werden. Das Ziel dieser Studie ist die Entwicklung eines ELISA
(enzyme-linked immunosorbant assay) als Hilfsmittel bei der Diagnostik von caniner Dermatophytose. Das verwandte Antigen war ein Ganzkrperextrakt von einem Isolat von M. canis, welches auf einem Flssigmedium
mit infizierten Haaren einer Katze mit fleckiger Alopezie kultiviert wurde. Um die Leistungsfhigkeit des ELISA
zu beurteilen, wurden die Seren von 18 Hunden mit Dermatophytose durch M. canis (Gruppe A, n = 18), 20
Hunden mit nicht-dermatophytischen Hauterkrankungen und 22 gesunden Hunden (Gruppe B, n = 42) getestet.Vier
weitere Tiere wurden getestet, drei mit Dermatophytose durch M.gypseum und eines durch T. mentagrophytes.
Ein signifikanter Unterschied (P < 0.01, Wilcox-Test, w = 364) bestand zwischen dem Ig-G-spezifischen Gehalt
von Seren von krzlich mit M. canis infizierten Hunden (Infektion < 15 Tage) und Kontrollen (obwohl drei Hunde
in diesem Stadium negative Titer hatten), whrend ein hoch signifikanter Unterschied zwischen den Kontrollen
und den Hunden mit einer Infektion von lngerer Dauer (> 30 Tage) festgestellt wurde. Alle Hunde hatten zu
diesem Zeitpunkt positive Titer. Eine hochgradig signifikante Korrelation (P < 0.001, Spearman-Test, rho =
0.86) wurde zwischen der Dauer der Infektion und der IgG Konzentration festgestellt. Der Test hatte gute
Sensitivitt (83.3%) und hohe Spezifitt (95.2%), aber einige Hunde behielten positive Titer nach Elimination der
Infektion. Die Sensitivitt ist hher als die direkte mikroskopische Haaruntersuchung und ist hnlich der einer
Pilzkultur mit DTM (Dermatophyten-Test-Medium).

2005 European Society of Veterinary Dermatology, Veterinary Dermatology, 16, 102107