Delvelopment And Application Of Latex Agglutination Method For Rapid Detection Of Canine Parvovirus

The disease that caused by Canine parvovirus (CPV) is one of the animal infectious doing serious damage to pets.It was first reported in China by Xun Hankun in1983. Although it is only30years since it was first detected in the world, but it has done very serious economical loss to pet breeding, owing to its high morbidity and mortality.CPV is a single-strand DNA virus, but the mutation rate is as high as the RNA virus, there have been three subtypes since it emerged in1977, named CPV-2a, CPV-2b, CPV-2c. It was devided to two types according to clinical signs:enteritis and myocarditis. But now myocarditis is rare, enteritis is comon.In pet hospital, imported colloidal gold Kit is used to detect canine parvovirus, but it is very expensive.This study aims to establish a latex agglutination assay,which is very easy,fast and cheap for rapid detecting of CPV.One of the students who have graduated from our laboratory had developed anti-CPV-2b monoclonal antibody.This study uses the McAb to sensitize the imported latex, then optimize the conditions for the sentization such as the amount of antibody and incubation time. The latex sensitized by the CPV McAb is stable and enjoy good specificity and high sensitivity.Finally latex agglutination method for detecting canine parvovirus was established and it was used to detect clinical samples.This study collected about150samples from pet hospital in Wuhan, Hubei Province.91samples of the150samples inoculated F81cells. After cytopathogentic effect was detected by transmission electron microscopic, then extracting the CPV DNA template, PCR amplification using the primers designed to get the583bp products. If the sample can amplify583bp fragment then it was senqenced by Aoke Company in Beijing. Analysis the results of sequencing,it shows that9of the91samples are CPV-2b(9.9%),72of91samples are CPV-2a(79.1%).The latex agglutination method was used to detect the samples comparing with PCR and colloidal gold methods.91samples were positive detecting by colloidal gold method, but the positive rate are only88%and84%, using PCR and latex agglutination method respectively. Moreover another57samples were detected both positive (48) or negative (4) by latex agglutination and colloidal gold method. They share the correlation of91.23%.Above all, this study establish a latex agglutination method for canine parvovirus rapid detection and it was applified in150clinical samples and the result is in accordance with PCR and Colloidal gold detection methods.It shares the correlation of91.23%with colloidal gold detection method. The molecular epidemiology invest shows that CPV-2a is the main subtype circulating in Wuhan. CPV-2b subtype was also isolated, but CPV-2c subtype was not isolated in Wuhan. This study lay a solid foundation for the prevention and treatment of canine parvovirus and provide a method for clinical detection of canine parvovirus.