Objective—To compare 5 methods of preparation of
RNA from feline urine samples for use in a feline calicivirus
(FCV), p30 gene-based, real-time reverse-transcriptase
polymerase chain reaction (RT-PCR) assay.
Sample Population—Urine and blood samples from
6 specific-pathogen-free cats.
Procedures—Aliquots of each urine sample (unmodified,
centrifuged, or mixed with whole or hemolyzed
blood) were spiked with FCV and serially diluted in
urine. Serial dilutions of FCV in tissue culture medium
were used as positive controls. Viral RNA was prepared
via dilution and thermal inactivation (DT
method), polyethylene glycol precipitation (PEG
method), isolation with oligo(dT)25-coated magnetic
beads (dTMB method), or extraction by use of 2 silica
gel–based columns (RN or QA method). Lower detection
limits and mean RT-PCR threshold cycle (Ct) values
associated with each RNA preparation method
and sample type were compared.
Results—Because DT-prepared samples yielded negative
results via RT-PCR assay, this method was not evaluated.
Lower detection limits (TCID50/sample) for the
assay in urine were 1,950, 104, 11, and 7 for PEG-,
dTMB-, RN-, and QA-prepared samples, respectively. For
RN and QA preparations, Ct values were similar and significantly
lower than those for dTMB and PEG preparations.
Overall, urine modifications did not affect FCV RNA
detection in dTMB-, QA-, and RN-prepared samples.
Conclusions and Clinical Relevance—Of the methods
evaluated, the RN and QA methods of RNA
preparation were most appropriate for the FCV RTPCR
assay. An RT-PCR assay optimized for detection
of FCV in feline urine may aid investigations of FCVinduced
urinary tract diseases in cats. (Am J Vet Res
Objective—To determine clinical, histologic, and immunohistochemical findings for dogs with wart-like lesions involving the paw pads.
Design—Retrospective case series.
Animals—24 dogs (18 Greyhounds and 6 dogs of other breeds).
Procedures—Medical records were reviewed for information on signalment, physical examination findings, concurrent disease processes, location of all lesions, and, when available, results of histologic examination of biopsy specimens. Available biopsy specimens (n = 11) were submitted for immunohistochemical staining and a PCR assay to identify viral inclusion bodies.
Results—In Greyhounds, most lesions involved the pads of the third and fourth digits, had a consistent histologic appearance without evidence of inflammation, were negative for papillomavirus, and had an unsatisfactory response to treatment. In other breeds, lesions often involved the pads of non–weight-bearing digits, had histologic evidence of inflammation, were positive for papillomavirus, and responded to surgical treatment.
Conclusions and Clinical Relevance—Results suggested that wart-like lesions involving the paw pads of Greyhounds were a distinct clinical entity with features resembling porokeratosis plantaris discreta in humans. In Greyhounds, these lesions were not associated with an underlying viral etiology and, therefore, should not be considered plantar warts. Alternative treatments should be investigated because current treatments were generally unsuccessful in Greyhounds. Wart-like lesions of the paw pads in other breeds were often associated with papillomavirus, and surgical excision appeared curative.
Objective—To evaluate 2 rapid, patient-side assays
for detection of Cryptosporidium parvum in feces
from neonatal calves with diarrhea.
Design—Diagnostic test evaluation.
Sample Population—Fecal samples from 96 neonatal
(1 to 30 days old) calves with diarrhea.
Procedure—Results of the rapid assays were compared
with results of microscopic examination of fecal smears
that had been stained with diamant fuchsin stain.
Results—One of the rapid assays correctly identified
56 of 62 (90%) fecal samples positive for C parvum
oocysts and 33 of 34 (97%) fecal samples negative
for oocysts. The other assay correctly identified 53 of
62 (85%) fecal samples positive for oocysts and 33 of
34 (97%) fecal samples negative for oocysts.
Conclusions and Clinical Relevance—Results suggest
that these 2 rapid assays are accurate when
used to detect C parvum in fecal samples from
neonatal calves with diarrhea. ( J Am Vet Med Assoc 2004;225:1090–1092)
Case Description—A 5-month-old captive female striped skunk (Mephitis mephitis) was evaluated because of lethargy, signs of depression, azotemia, and erythema of the skin around the eyes.
Clinical Findings—Antemortem diagnostic tests revealed renal disease but failed to identify an etiologic agent. A diagnosis of severe nonsuppurative interstitial nephritis was made on the basis of results of histologic examination of renal biopsy specimens.
Treatment and Outcome—The skunk was administered isotonic fluids SC daily and later every other day because of the handling-related stress. Because of the skunk's deteriorating condition, it was euthanized after 24 days of supportive care. Aleutian disease was diagnosed on the basis of positive results of a PCR assay that targeted the DNA from Aleutian disease virus (ADV); positive results for ADV were also obtained by use of plasma counterimmunoelectrophoresis and an ELISA. Genetic sequencing of the 365-base pair PCR product revealed 90% sequence identity with mink ADV.
Clinical Relevance—In the skunk of this report, infection with a skunk-specific parvovirus resulted in clinical signs and pathologic changes similar to those associated with ADV infection in mink. For skunks with signs of renal failure, differential diagnoses should include parvovirus infection. In confirmed cases of infection with this ADV-like virus, appropriate quarantine and biosecurity measures should be in place to prevent spread to other susceptible animals within a zoological collection.