One-step immunochromatography assay kit for detecting antibodies to canine parvovirus

Abstract

This study was performed to determine the feasibility of using whole serum to detect antibodies to canine parvovirus (CPV) under nonlaboratory conditions and to evaluate the performance characteristics of an immunochromatography assay kit. Precise detection of levels of antibody against CPV in puppies can be used to determine a vaccination schedule, because maternal antibodies frequently result in the failure of protective vaccination, and can also be used to determine the antibody levels of infected puppies. Several methods for the titration of CPV antibodies have been reported, including the hemagglutination inhibition (HI) assay, which is considered the "gold standard." These methods, however, require intricate and time-consuming procedures. In this study, a total of 386 serum specimens were tested. Compared to the HI assay, the rapid assay had a 97.1% sensitivity and a 76.6% specificity (with a cutoff HI titer of 1:80). This single-step assay could be performed rapidly and easily without special equipment. The kit provides a reliable method for detection of anti-CPV antibody where laboratory support and personnel are limited.

FIG. 1.

Diagram of the test strip for the detection of anti-canine parvovirus antibody. Serum is added to the sample pad where serum antibodies can interact with CPV. Addition of buffer enables the complex to migrate along the test strip where gold-conjugated antibodies are captured by the immobilized anti-porcine or anti-canine IgG.

FIG. 2.

Western blotting with CPV MAb 4c3. CPV was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with CPV MAb 4c3. The CPV MAb 4c3 bound to the 64-kDa protein of CPV. M, molecular markers.

FIG. 3.

The intensity of the color developed at the T line (T) correlated with the HI assay-determined titer of the reference serum sample shown on each test strip.