A previously described monoclonal antibody (MAb)-based competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was modified to optimize performance, and the assay was validated in various defined cattle populations for detection of serum antibody to Neospora caninum, a major cause of bovine abortion. Modifications to the cELISA included capturing native N. caninum antigen with a parasite-specific MAb (MAb 5B6-25) and directly conjugating the competitor MAb (MAb 4A4-2), with both MAbs binding different epitopes of a conserved, immunodominant 65-kDa tachyzoite surface antigen. The assay was validated using three serum sets, a "gold standard" set of 184 cow sera defined by fetal histopathology and N. caninum immunohistochemistry and by maternal N. caninum indirect fluorescence assay (IFA) at a 1:200 serum dilution, a relative standard set of 330 cow sera defined by IFA alone, and a set of 4,323 cow sera of unknown N. caninum status. A test cutoff of 30% inhibition was identified. The diagnostic sensitivity was 97.6%, and diagnostic specificity was 98.6% for the gold standard abortion-defined sera. The diagnostic sensitivity was 96.4%, and diagnostic specificity was 96.8% for the relative standard IFA-defined sera. Testing of the 4,323 bovine sera of unknown N. caninum status revealed a distinct bimodal distribution and steep sigmoid frequency curve with only 1.8% of samples within 5% of the test cutoff, indicating a sharp discrimination between test-positive and test-negative samples. In summary, the modified N. caninum cELISA provided a simple, rapid, and versatile method to accurately identify N. caninum infection status in cattle using a single cutoff value.
A high number of reported canine leptospirosis cases occurred in Washington State from 2004 to 2006. This prompted a serosurvey of healthy dogs from around the state to determine the distribution of exposure risk and to provide insight into serovar epidemiology in the region. In addition, a convenience sample of sera from injured raccoons was also tested, and clinical serological data from the Washington Animal Disease Diagnostic Laboratory were examined. The proportion of dogs with an antibody titre (>or=1:100) to any serovar was 27/158 (17.1%, 95% CI 11.6-23.9), and that proportion among raccoons was 22/115 (19.1%, 95% CI 12.4-27.5) suggesting that the potential for exposure in Washington state is not uncommon. The most frequently detected serovars in healthy dogs were Autumnalis, Icterohemorrhagiae and Canicola, in clinical canine samples Autumnalis, Bratislava and Pomona were more frequent and in sick or injured raccoons Autumnalis, and Pomona were most frequently detected. Clinical canine serology demonstrated a late summer-fall seasonality that was consistent with other reports. An outbreak of canine leptospirosis occurred during 2004-2006 and was located primarily in western Washington counties, as were three reported human cases in 2005. Canine leptospirosis surveillance is an important tool for detecting human risk of exposure and may provide insights into which serovars are currently of clinical importance.
The sensitivity of the agar gel immunodiffusion (AGID) test for the detection of antibody to caprine arthritis-encephalitis virus (CAEV) was investigated with CAEV or ovine progressive pneumonia virus (OPPV) as the source of antigen. A total of218 goat serum specimens were tested for anti-CAEV antibody by AG!D and immunoprecipitation of [33Sjmethionine-labeled CAEV. In comparison with that of immunoprecipitation, the
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.