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BlackwellArticle
Original
Science, Ltd

Evaluation of an enzyme-linked immunosorbant assay

(ELISA) for the serological diagnosis
of sarcoptic mange in dogs
KIMBERLY S. LOWER,* LINDA M. MEDLEAU,* KEITH HNILICA
and BEAT BIGLER
*University of Georgia, College of Veterinary Medicine, Department of Small Animal Medicine,
Athens, GA 30602, USA
University of Tennessee, College of Veterinary Medicine, Department of. Small Animal Clinical Sciences,
Knoxville, TN 37901-1071, USA
Labor Laupeneck/imovet bg, Laupenstrasse 33, CH-3008 Bern, Switzerland
(Received 1 November 2000; accepted 22 January 2001 )

Abstract Canine scabies is a challenging disease to diagnose because sarcoptic mites are hard to find on skin
scrapings. The purpose of this study was to evaluate a serologic enzyme-linked immunosorbant assay (ELISA)
as an aid in the diagnosis of canine scabies. In addition, serum samples were obtained post treatment to determine the duration and persistence of circulating scabies antibodies after resolution of natural infection. Nineteen
dogs diagnosed with sarcoptic mange and 38 control dogs were tested. Sixteen scabies-infested dogs showed
positive pretreatment ELISA results (84.2% sensitivity). Thirty-four control dogs showed negative ELISA results
(89.5% specificity). In the 11 scabies dogs from which multiple post treatment serum samples were obtained,
detectable antibodies were not present 1 month after treatment in four cases, but were present for 14.5 months
post treatment in seven dogs. Our results suggest that this scabies ELISA test is useful in the diagnosis of
canine scabies.
Keywords: dog, ELISA, Sarcoptes scabiei, scabies, serodiagnosis.

INTRODUCTION
Canine scabies is a parasitic skin disease caused by the
mite Sarcoptes scabiei var. canis. The mite is easily
spread by direct contact and so is very contagious.
Clinical signs of scabies infestation can be variable. The
classic presentation is an intensely pruritic dog with
alopecia and crusting involving the ear pinnal margins
and pressure points, but some dogs have the more
vague presentation of a generally pruritic dog with few
or no skin lesions.1,2 Scabies can, therefore, closely
mimic other causes of pruritus such as atopy, food
allergy and flea allergy. The diagnosis of scabies can be
challenging, as mites are found in only 2050% of skin
scrapings from infested dogs.13 Scabies-suspect dogs
are often treated empirically either topically or systemically with acaricidal agents, and a positive response to
treatment is viewed as an indirect way of diagnosing
the disease. Because allergic skin disease has clinical
signs similar to scabies, many pruritic dogs are treated
empirically with acaricides either prior to, or in con-

Correspondence: Kimberly Lower, Dermatology Clinic for

Animals, 1580 E. Whitten St. Chandter, AZ 85225, USA. E-mail:
klderm@msn.com
2001 Blackwell Science Ltd

junction with, investigation for allergies. This increases

the time and money spent on the diagnosis of allergic
dogs, and exposes them unnecessarily to potentially
toxic medications. Conversely, some scabies-infested
dogs that do not have the classical presentation are
erroneously diagnosed and treated for allergies at great
expense and animal discomfort.
Infestation with scabies elicits a measurable antibody response in mammals.311,14 17,19,22 Skin biopsy
specimens from scabies-infested skin show increased
immunoglobulin-secreting cells,12 as well as locally
increased immunoglobulins and complement.13
Enzyme-linked immunosorbant assays (ELISA) for
scabies antibodies have been developed for pigs and
dogs.3,10,1416 Diagnosis of canine scabies in Europe
has been enhanced by the development of a scabies
antibody ELISA test, but this test is unavailable outside Sweden, although serum samples can be sent to
the Swedish laboratory for analysis. A similar scabies
ELISA test that is commercially available outside
Europe has recently been developed in Switzerland.
The purpose of this study was to evaluate the
Swiss scabies ELISA test in the diagnosis of scabies in
American dogs. Post treatment sera samples were also
evaluated in dogs with scabies to determine duration
of persistence of scabies antibodies after resolution of
natural infection.
315

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K. S. Lower et al.

MATERIALS AND METHODS

Dog sera
Serum samples were collected from 19 dogs (five 8week-old puppies, four 12-week-old puppies and 10
dogs > 6 months of age) with natural scabies infestation, verified by positive identification of Sarcoptes
scabiei mites and/or ova on skin scrapings. All infested
dogs were treated successfully with scabicidal therapy
(systemic ivermectin, topical insecticidal dips, selamectin or fipronil spray). Pretreatment serum samples
were obtained from all 19 dogs. In 11 of the dogs, 1
4 monthly post treatment serum samples were also
obtained.
Thirty-eight negative control serum samples were
collected from dogs with no history of skin disease
(n = 10), and from dogs with skin diseases other than
scabies (n = 28). In dogs with skin disease, scabies was
ruled out via negative skin scrapings, lack of complete
response to trial scabicidal therapy, and resolution of
clinical signs with nonscabicidal treatments. This
included 18 atopic dogs, 5 dogs with demodicosis, 3
dogs with food allergy verified by recurrence of pruritus with diet challenge, and 2 dogs with miscellaneous
disorders (lick granuloma and pyoderma). Fifteen of
the 18 atopic dogs had positive intradermal (n = 14)
or serological (n = 1) reactions to house dust and/or
house dust mite; the remaining three atopic dogs
showed reactivity to pollens, insects and/or moulds,
but not to house dust or house dust mite.
All serum samples were frozen immediately and
stored at 20 C until analysis; samples were sent as a
group to the Swiss laboratory (laboratory name:
imovet bg) for analysis.
Serologic testing
Sera were analysed with an indirect ELISA (test name:
Imovet sarcoptes) based on S. scabiei var. vulpes using
a polyclonal goat antidog immunoglobulin (Ig)G
conjugated with alkaline phosphatase (Kirkegaard and
Perry Co., USA). Serum from a Swiss dog with sarcoptic
mange verified by skin scraping was used as a positive control, and serum from a Swiss dog raised under
parasite-free conditions was used as a negative control.
Optical densities (OD) were read 1 h after substrate
addition. The raw data of the measured OD was then
expressed as a percentage of the positive control in a
ratio correcting for the measured OD of the negative
control: (OD sample OD negative control)/(OD positive control OD negative control). In accordance
with laboratory reference data, converted OD values
> 20% were considered positive, and converted OD
values < 20% were considered negative.
Statistical analysis
Results of scabies tests were correlated to case diagnosis
to establish sensitivity and specificity results and analysed
using a Fishers exact 2 2 contingency table which
was performed using the software program 3.30 A P-value < 0.05 was considered significant.
2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 315 320

Table 1. Scabies ELISA results: cases and controls

Positive serology
Negative serology
Total

Scabies
cases

All
controls

16
3
19

4
34
38

P-value < 0.0001, Fishers exact test.

RESULTS
Sera from dogs with scabies
Positive pretreatment ELISA test results were obtained
for 16 of 19 scabies-infested dogs (84.2% sensitivity,
Table 1). The mean converted OD value obtained for
scabies-infected dogs was 53.5% (range SD: 5.7
101.3 30.49%, Fig. 1). Two false-negative results
(converted OD values 5.7 and 14.7%) were from 8-weekold puppies which had symptoms for an unknown
period prior to sampling. One of these puppies had
monthly sequential samples obtained for 3 months
post treatment; all samples showed negative ELISA
results. The third falsely negative result (converted
OD, 17.0%) was obtained from a 5-year-old dog with
a 6-month history of pruritus treated by daily corticosteroid administration (prednisone 12 mg kg1).
Although the pretreatment titre on this dog was initially negative, sequential results were positive at 1
(converted OD, 62.0%) and 2 months post treatment
(converted OD, 33.5%).
In the other nine affected dogs from which multiple
samples were obtained, significantly detectable antibodies were not present 1 month after treatment in
three cases (one 8-week-old puppy, one 12-week-old
puppy and one dog > 6 months of age), but persisted
for 1 month post treatment in one dog (a 12-week-old
puppy) and for 24.5 months post treatment in five
dogs (one 8-week-old puppy, two 12-week-old puppies
and two dogs > 6 months of age) (data not shown, but
available upon request).
Negative control sera
Of 38 control dogs, 34 negative ELISA results were
obtained (89.5% specificity, Table 1). The mean converted OD value for control dogs was 10.0%
(range SD: 048.5 9.61%, Fig. 1). Two positive
results (converted OD values, 20.0 and 31.0%) were
obtained from normal dogs with no history of skin
disease or scabies exposure. No positive results were
obtained from the Demodex dogs. Two positive results
were obtained from atopic dogs with positive intradermal reactions to house dust and/or house dust mite
(Table 2). Of these two dogs, one (converted OD,
29.3%) responded well to allergy hyposensitization
and was never treated for scabies, and the other
(converted OD, 48.5%) was trial treated for scabies
twice (parenteral ivermectin, amitraz dips) with minimal effect; immunotherapy was initiated for this
patient.

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Scabies and ELISA

317

Figure 1. Mean scabies ELISA results.

Table 2. Distribution of controls

Positive serology
Negative serology
Total

Statistical analysis
The Fishers exact test revealed a P-value < 0.0001,
confirming a significant association between ELISA
result and case status.

DISCUSSION
Scabies infestation causes the production of measurable antibodies in infested host species.311,1417,19,22 This
study demonstrates that this ELISA test is accurate
in the detection of antibodies to S. scabiei in dogs,
with a sensitivity of 84.2% and specificity of 89.5%.
This is comparable with a recent study carried out in
the UK using the Swedish scabies ELISA test which
analysed sera samples from 13 normal dogs, 12 atopic
dogs and 12 documented scabies dogs (83% sensitivity and 92% specificity).29 In our study, microscopic
identification of scabies mites and/or ova was used to
diagnose infestation, in contrast to other studies in
which clinical signs and response to treatment were used
to confirm the diagnosis in the majority of affected
animals.3,10,11,1416 This may account for the lower
sensitivity and specificity values obtained in this, and
the UK study, compared with the original Swedish
study (sensitivity 92%, specificity 96%).3 Although
diagnosis of scabies by response to treatment is more

Normal /
nonskin
disease
2
8
10

Atopy

Food
allergy

Demodex

Other
(lickgranuloma,
pyoderma)

2
16
18

0
3
3

0
5
5

0
2
2

convenient and clinically realistic, as mites are found in

< 50% of skin scrapes, interpretation of results could
be complicated by occult infestation of suspect animals
with nonscabies parasites (which would also respond
clinically to parasiticidal therapy), or by the occasional
treatment failure of scabies-infested patients.13
The ELISA utilized in this study was based on the
fox variant of S. scabiei. Although use of the canine
variant would be ideal, infested dogs typically have few
mites.1 Large numbers of vulpes variant mites can be
obtained for the manufacture of test reagents. Canine
scabies cannot be propagated in vitro, and can only be
propagated in vivo in laboratory rabbits.8,9 Prior studies
have demonstrated that subspecies of S. scabiei share
common antigens which are recognized by antibodies
of different host species.7,17,19 The various strains are
morphologically alike, and host specificity is likely due
to physiological differences.17,19 ELISA tests based on
S. scabiei var. vulpes have previously been used successfully to detect sarcoptic antibodies in pigs infected with
S. scabiei var. suis and in dogs infected with S. scabiei
var. canis.3,10,11,1416
The sensitivity of this test was 84.2%, due to three
dogs (two puppies and one adult) that had no measurable antibodies in spite of demonstrable mites on skin
scrapings. One potential reason for this incongruity is
lack of sufficient time for seroconversion to occur.
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Dogs infested experimentally with scabies seroconverted 35 weeks post infection, or 13 weeks after the
onset of clinical signs.4 Sampling of the falsely negative
puppies may thus have occurred during the 13 week
window between onset of pruritus and seroconversion.
Another possible cause for the discordant results may
have been the young age (8 weeks) and immature
immune system of the puppies. A similar finding of
false-negative results in 78-week-old piglets infected
with S. scabiei var. suis was theorized to have been due
to inadequate production of specific antibodies, possibly secondary to interference from high maternal antibodies.11 A potential cause for the falsely negative titre
in the adult dog may have been prior treatment with
corticosteroids, which inhibit T- and B-cell function
and antibody production.27 This dog had clinical disease for 6 months and had been treated with high doses
of steroids. The initial ELISA result was negative, but
sequential samples became positive at 1 and 2 months
post treatment, after steroids had been discontinued.
Seroconversion post treatment in this dog may have
also been related to an increased immune reaction to
antigens released by dead and decomposing mites.18
Lastly, false-negative reactions can be due to laboratory error, such as prolonged storage of nonfrozen sera
samples, samples containing sodium azide, allowing
the chromagen solution or treated plates to be exposed
to light, or allowing plates to dry out between the different stages of the test. Elimination of these latter
potential causes is done by careful sample collection
and storage and by adherence to correct testing protocol. To rule out laboratory error as the cause for the
falsely negative results, duplicate serum samples from
the dogs in question were later resubmitted for repeat
testing and found to have the same results.
Measurable antibodies to scabies mites were found
14.5 months post treatment of natural infestation in
7 of 11 dogs in which antibody levels were followed.
This result is similar to previous studies of experimental
scabies infestation.4,6,8 This finding limits the use of
the scabies ELISA test in the evaluation of scabicidal
treatment efficacy in dogs which have an incomplete
clinical response to therapy.
The specificity of this test was 89.5%, because of four
positive results in negative control dogs which did not
have demonstrable scabies mites on skin scrapings, did
not respond completely to empirical scabicidal therapy
and did respond to noninsecticidal therapy. Inclusion
of more nonpruritic dogs in the control group may
have increased this value, however, use of pruritic
controls gives a more realistic measurement as in clinical use this test would not be performed in nonpruritic
dogs. The most likely reason for false positive results
would be prior exposure to scabies or concurrent
scabies infection in an allergic patient. Prior exposure
is strongly possible as this study shows that in natural
scabies infestation detectable antibodies may persist
for at least 4.5 months after successful treatment. Also,
as animals with allergic skin disease are frequently at
veterinary hospitals and grooming establishments,
2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 315 320

their risk for exposure to the mite may be increased.

One of the seropositive control dogs (an atopic dog
that reacted to house dust mite) was never treated for
scabies but responded clinically to allergy immunotherapy. In addition to prior exposure to scabies, there
is the possibility that this dog was truly infested and
that immunotherapy with extracts of house dust mite
containing antigens cross-reactive with S. scabiei may
have elicited protective immunity as demonstrated for
rabbits, causing clearance of the mites and resolution
of clinical signs.25 Less likely reasons for falsely positive
results include laboratory error, such as improper
washing of the plate or contamination of the diluent or
reagents. To rule out laboratory error as the cause for
the falsely positive results, duplicate serum samples
from the dogs in question were later resubmitted for
repeat testing and found to have the same results.
Alternatively, falsely positive serologic results to
S. scabiei may be due to cross-reactivity with antibodies to house dust mite (Dermatophagoides farinae
and D. pteronyssinus), which have been shown in vitro
and in vivo to share antigens with S. scabiei.18 21,23 26
However, only 2 of 15 atopic dogs (13.3%) that reacted
to house dust and/or house dust mite showed positive
results on the scabies ELISA. These results are similar
to a recent study investigating the Swedish scabies
ELISA which found no positive scabies results in 12
house dust mite-reactive atopic dogs.29 Serum antibodies of human atopic patients sensitive to house dust
mite bind to multiple S. scabiei antigens, and atopic
patients with no history of scabies exposure show
positive skin test reactivity to D. pteronyssinus and
S. scabiei.22,24 Conversely, dogs infested with scabies
showed increased antibody formation not only to
scabies, but also to D. farinae and D. pteronyssinus.18,28
There is evidence that prior exposure to scabies may
predispose affected individuals to the development of
clinical sensitivity to house dust mite, and response to
such allergens may explain the persistence of symptoms in some cases after treatment of scabies.19,23 In
addition, individuals sensitive to dust mites may be
more prone to develop scabies and to have more severe
clinical signs than nonallergic individuals.20,21 Although
only two of our house dust mite-sensitive atopic dogs
were positive on the scabies ELISA, these factors may
make interpretation of scabies serology more complicated in dust mite-sensitive patients.
In summary, although interpretation of positive results
may be occasionally problematic in atopic patients
sensitive to house dust mites, this study illustrates the
potential usefulness of a commercially available ELISA
in the diagnosis of canine scabies. The specificity and
sensitivity are acceptable and similar to other scabies
ELISA tests.3,10,11,1416,29 The successful treatment of
dogs with pruritic skin diseases must be based on a correct diagnosis which can differentiate between scabiesinfected and noninfected dogs. Diagnostically, this
ELISA is more accurate than skin scrapes, which are
informative in < 50% of cases,1 3 making serology
helpful in the diagnosis and therapy of pruritic dogs.

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Scabies and ELISA

ACKNOWLEDGEMENTS
This work should be attributed to the University of
Georgia, College of Veterinary Medicine, Department
of Small Animal Medicine. This study was self-funded.

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Rsum La gale est une maladie difficile diagnostiquer chez le chien, car les Sarcoptes sont difficiles trouver
sur les raclages cutans. Le but de cette tude tait dvaluer un test ELISA comme aide au diagnostic de la gale
sarcoptique chez le chien. En outre, des prlvements sriques ont t obtenus aprs traitement pour dterminer
la dure et la persistance des anticorps circulants aprs gurison de linfection naturelle. Dix neuf chiens prsentant une gale sarcoptique et 38 chiens contrles ont t tests. Seize chiens infects avaient un test ELISA positif
avant traitement (sensibilit de 84.2%). Trente quatre chiens contrles avaient un test ngatif (spcificit de
89.5%). Chez les 11 chiens galeux pour lesquels des prlvements ont t obtenus aprs traitement, aucun anticorps na t dtect 1 mois aprs le traitement pour 4 cas, mais pour les sept autres chiens des anticorps taient
2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 315320

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K. S. Lower et al.
encore prsents 1 4.5 mois aprs traitement. Nos rsultats suggrent que ce test ELISA est utile dans le diagnostic
de la gale sarcoptique chez le chien.
Resumen La sarna sarcptica en el perro es una enfermedad de diagnstico difcil debido a la dificultad en
encontrar caros en raspados cutneos. El objetivo de este estudio fue evaluar una prueba de inmunoabsorbente
ligado a enzima (ELISA) como ayuda en el diagnstico de la sarna sarcptica canina. Adems, se obtuvieron
muestras de suero despus del tratamiento para determinar la duracin y la persistencia de anticuerpos circulantes de sarna sarcptica despus de la resolucin de la infeccin natural. Diecinueve perros con un diagnstico
de sarna sarcptica y 38 perros control fueron testados. Diecisis perros infestados mostraron resultados ELISA
positivos antes del tratamiento (84.2% de sensibilidad). Treinta y cuatro perros control mostraron resultados
ELISA negativos (89.5% especificidad). En los 11 perros con sarna sarcptica, de los cuales se obtuvieron
mltiples muestras de suero despus del tratamiento, no se detectaron anticuerpos 1 mes despus del tratamiento
en cuatro casos, pero estuvieron presentes durante 1 a 4.5 meses despus del tratamiento en siete perros. Nuestros
resultados sugieren que la prueba de ELISA para sarna sarcptica es til en el diagnstico de esta enfermedad.
Zusammenfassung Sarkoptesrude beim Hund ist eine diagnostische Herausforderung, weil Sarkoptesmilben
auf Hautgeschabseln schwer zu finden sind. Der Zweck dieser Studie war es, einen serologischen Enzym-linked
immunosorbent Assay (ELISA) als diagnostische Hilfe bei der Sarkoptesrude des Hundes zu bewerten.
Zustzlich wurden nach der Behandlung Serumproben genommen, um die Zeitdauer und Persistenz der zirkulierenden Serumantikrper nach Resolution der natrlichen Infektion zu bestimmen. Neunzehn mit Sarkoptesrude diagnostizierte Hunde und 38 Kontrollhunde wurden getestet. Bei 16 mit Sarkoptes befallenen Hunden
war der ELISA positiv (84.2% Sensitivitt). Bei 34 Kontrollhunden war der ELISA negativ (89.5% Spezifizitt).
Bei den 11 Hunden, bei denen mehrere Serumproben nach der Behandlung entnommen wurden, waren
Antikrper in 4 Fllen einen Monat nach der Behandlung nicht mehr nachweisbar, wurden aber bei 7 Hunden
nach 14.5 Monaten immer noch nachgewiesen. Unsere Ergebnisse deuten darauf hin, dass dieser Sarkoptes
ELISA Test bei der Diagnose von Sarkoptesrude beim Hund ntzlich ist.

2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 315 320