Impact of heat treatment on Dirofilaria immitis antigen detection in shelter dogs.

BACKGROUND: The diagnosis and management of canine heartworm disease is a growing concern for shelter veterinarians. Although the accuracy of commercial antigen test kits has been widely studied, recent reports have renewed interest in antigen blocking as a causative factor for false "no antigen detected" results. The objectives of this study were to determine the prevalence of false "no antigen detected" results in adult dogs entering shelters in northern, southern, and western regions of the country and to identify historical and clinical risk factors for such results. METHODS: Serum samples were evaluated for Dirofilaria immitis antigen using a commercially available point-of-care ELISA; samples in which no antigen was detected underwent a heat treatment protocol and repeat antigen testing. Whole blood samples underwent Knott testing to identify the presence of microfilariae. Historical and clinical findings were analyzed using exact logistic regression. RESULTS: A total of 616 samples were analyzed. Overall prevalence of positive antigen test results (prior to heat treatment) was 7.3% and frequency of false "no antigen detected" results due to antigen blocking (ie, samples with no antigen detected prior to heat treatment and positive after heat treatment) was 5.2%. Among dogs that had no detectable antigen on the initial tests, dogs that had microfilariae detected via modified Knott testing (OR = 32.30, p-value = 0.013) and dogs that previously received a heartworm preventive (OR = 3.81, p-value = 0.016) had greater odds of antigen blocking than dogs without these factors. Among dogs that were heartworm positive, those without microfilariae detected had greater odds of antigen blocking than dogs with this factor (OR = 11.84, p-value = 0.0005). Geographic region of origin was significantly associated with occurrence of antigen blocking (p = 0.0036); however, blocking occurred in all regions sizably contributing to heartworm diagnoses. Of the 74 dogs found to be infected with heartworms in this study, 39.2% (29) had no detectable antigen prior to heat treatment. CONCLUSIONS: Heat treatment of serum samples should be considered to improve diagnostic test accuracy, particularly in dogs that reportedly received a heartworm preventive prior to antigen testing regardless of region of origin.

Prevalence of methicillin-resistant staphylococci in northern Colorado shelter animals.

Methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP) have been recognized as significant pathogens in veterinary medicine. There have been documented cases of MRSA infection and colonization in veterinary critical care units, in veterinary personnel, and in equine and feline patients. To date, there have been no studies examining the prevalence of MRSA or MRSP colonization of cats and dogs in animal shelters in the United States. The purpose of the current study was to determine the prevalence of MRSA and MRSP in cats and dogs in a northern Colorado animal shelter. Samples were collected from 200 cats and 200 dogs in an open admission shelter. Each species was divided into 2 smaller groups: 100 dogs or cats housed in the stray ward and 100 dogs or cats housed in the adoption area. Samples were evaluated for the prevalence of MRSA or MRSP, which was verified through aerobic culture and Kirby-Bauer agar disc diffusion to confirm antimicrobial sensitivity. Results revealed MRSA in 0.5% of cat samples, MRSA in 0.5% of dog samples, and MRSP in 3% of dog samples. These results are consistent with previously published prevalence rates for these 2 organisms in non-shelter populations of dogs and cats, indicating that cats and dogs from this Colorado shelter do not appear to pose any greater risk to the public than do cats and dogs in the general pet population.

Validation of an anti-PA-ELISA for the potency testing of anthrax vaccine in mice.

The potency test for the anthrax vaccine currently licensed for human use in the United States (Anthrax Vaccine Adsorbed) involves the protection of actively immunized guinea pigs from a lethal challenge with a virulent strain of Bacillus anthracis. Lethal challenge tests entail the use of specialized containment facilities for the safe and secure handling of the challenge strain. This potential difficulty, plus humane considerations, have prompted us to investigate non-lethal, alternative immunogenicity assays that could be considered as potency tests not only for the current vaccine, but also for vaccines under development. Immunogenicity tests will require suitable measurement of an antibody response to relevant antigens, by methods such as enzyme linked immunosorbent assay (ELISA) or a toxin neutralization assay. Any assay chosen for this purpose should be adequately validated and reproducible by other laboratories. Validation of an analytical procedure requires the demonstration that the assay is suitable for its intended purpose. The objective of this work was to study the performance of an anti-PA-ELISA designed to assess the antibody response to anthrax vaccines in mice. Validation studies were performed according to the guidelines of the International Conference of Harmonization (ICH), and we have established the working range of the assay (37-1159 EU/mL) on the bases of the following parameters: linearity (20-1159 EU/mL; r2=0.99; p-value=0.21), accuracy (91-118% recovery), precision (< or =20%CV, repeatability; < or =9 and < or =21%CV, intermediate precision per day and per analyst, respectively), detection limit (5 EU/mL), and quantification limit (37 EU/mL). We believe that assay specificity and the above characteristics are adequate to allow this ELISA to be considered for use in a mouse immunogenicity (potency) test of anthrax vaccines, and for the standardization of reagents.