A polyclonal antibody-based antigen-capture ELISA (AC-ELISA) has been developed for detection of Canine parvovirus (CPV) antigens in faecal samples of dogs. The assay uses rabbit anti-CPV polyclonal antibody as the capture antibody, guinea pig anti-CPV polyclonal antibody as tracing antibody and anti-guinea pig HRPO conjugate as the detection system. The optimum dilution of the capture antibody and the tracing antibody capable of detecting the CPV-2 antigens was found to be 1:1 600 and 1:400, respectively, in the check-board titration. In this study, a total of 152 samples (129 faecal samples and 23 cell culture supernatant) were tested both by AC-ELISA and by polymerase chain reaction (PCR). Of the samples tested, 69 and 78 samples were found positive by AC-ELISA and PCR, respectively. The AC-ELISA had relative sensitivity, relative specificity and accuracy of 88.4%, 100.0% and 91.4% respectively. The analytical sensitivity of AC-ELISA was estimated to be 102.8 TCID50/mL whereas PCR sensitivity was 100.8 TCID50/mL. The AC-ELISA is a simple, quick and reliable method for screening large numbers of faecal samples of dogs suspected of CPV infection.
Appel M J G, Cooper B J, Greisen H, et al. 1978. Status report: canine viral enteritis. J Am Vet Med Assoc, 173: 1516–1518.
Battilani M, Scagliarini A, Tisato E, et al. 2001. Analysis of canine parvovirus sequences from wolves and dogs isolated in Italy. J Gen Virol, 82: 1555–1560.
Bell C J, Finlay D A, Clarke H J, et al. 2002. Development of a sandwich ELISA and comparison with PCR for the detection of F11 and F165 fimbriated E. coli isolates from septicaemic disease in farm animals. Vet Microbiol, 85: 251–257.
Biswas S, Das P J, Ghosh S K, et al. 2006. Detection of canine parvovirus (CPV) DNA by polymerase chain reaction and its prevalence in dogs in and around Kolkata, West Bengal. Indian J Anim Sci, 76(4): 324–325.
Cho H S, Song J E, Park Y S, et al. 2004. Diagnosis of the canine parvovirus in faecal samples by in situ hybridization. Indian Vet J, 81: 855–859.
Decaro N, Elia G, Martella V, et al. 2005. A real-time PCR assay for rapid detection and quantitation of canine parvovirus type 2 in the feces of dogs. Vet Microbiol, 105: 19–28.
Decaro N, Desario C, Addie D D, et al. 2007. The study of molecular epidemiology of canine parvovirus, Europe. Emerg Infect Dis, 13: 1222–1224.
Deepa P M, Saseendrannath M R. 2000. Serological studies on canine parvoviral infection. Indian Vet J, 79: 643–644.
Hawkes R A, Kirkland P D, Sanders D A, et al. 2000. Laboratory and field studies of an antigen capture ELISA for Bluetongue virus. J Virol Meth, 85: 137–149.
Ho-Seong C, Jong K, Nam-Yong P. 2006. Detection of canine parvovirus in fecal sample using loop-mediated isothermal amplification. J Vet Diagn Invest, 18: 81–84
Joshi D V, Singh S P, Rao V D P, et al. 1998. Isolation of canine parvovirus from clinical cases of gastroenteritis. Indian Vet J, 75: 498–500.
Kumar P, Garg S K, Bandyopadhyay S K, et al. 2003. Haemagglutinating activity of canine parvovirus. Indian J Anim Sci, 73(2): 123–125.
Maree S, Paweska J T. 2005. Preparation of recombinant African horse sickness virus VP7 antigen via a simple method and validation of a VP7-based indirect ELISA for the detection of group-specific IgG antibodies in horse sera. J Virol Meth, 125: 55–65.
Martella V, Cavalli A, Pratelli A, et al. 2004. A canine parvovirus mutant is spreading in Italy. J Clin Microbiol, 42: 1333–1336.
Mochizuki M, SanGabriel M C, Nakatani H, et al. 1993. Comparison of polymerase chain reaction with virus isolation and haemagglutination assays for the detection of canine parvoviruses in fecal specimens. Res Vet Sci, 55: 60–63.
Mohan R, Nauriyal D C, Singh K B. 1993. Detection of canine parvovirus in faeces, using a parvovirus ELISA test kit. Indian Vet J, 70: 301–303.
Nakamura M, Tohya Y, Miyazawa T, et al. 2004. A novel antigenic variant of canine parvovirus from a Vietnamese dog. Arch Virol, 149: 2261–2269.
Nandi S, Chidri S, Kumar M. 2009. Molecular characterization and phylogenetic analysis of a canine parvovirus isolate in India. Vet Med Czech, 54(10): 483–490.
Nandi S, Chidri S, Kumar M, et al. 2010. Occurrence of canine parvovirus type 2c in the dogs with haemorrhagic enteritis in India. Res Vet Sci, 88: 169–171.
Narayanan S, Amit K, Chug P K, et al. 2001. Strain characterization of Indian isolates of canine parvovirus by PCR-RFLP. J Rem Vet Corp, 40: 67–74.
Parker J S, Murphy W J, Wang D, et al. 2001. Canine and feline parvoviruses can use human or feline transferrin receptors to bind, enter and infect cells. J Virol, 75: 3896–3902.
Pereira C A, Monezi T A, Mehnert D U, et al. 2000. Molecular characterization of canine parvovirus in Brazil by polymerase chain reaction assay. Vet Microbiol, 75: 127–133.
Phukan A, Deka D, Boro P K. 2004. Occurrence of canine parvovirus infection in and around Guwahati. Indian J Anim Sci, 74(4): 930–931.
Ramadass P, Khadher T G A. 1982. Diagnosis of canine parvovirus infection by agar gel precipitation test and fluorescent antibody technique. Cherion, 11: 323–326.
Reed L J, Muench H. 1938. A simple method of estimating fifty percent endpoints. American J Hyg, 27: 493–497.
Rimmelzwaan G F, Groen J, Juntti N, et al. 1987. Purification of infectious canine parvovirus from cell culture by affinity chromatography with MAbs. J Virol Meth, 15: 313–322.
Rimmelzwaan G F, Groen J, Boarst G H A, et al. 1991. The use of ELISA system for serology and antigen detection in parvovirus, coronavirus and rotavirus infection in dogs in the Netherland. Vet Microbiol, 26: 25–40.
Ruiz D E, Banez R. 1996. Comparison of sandwich ELISA with AGID and CIE for diagnosis of canine parvovirus. Vet Med, 13: 665–669.
Tabor A E, Patterson R M, Smith J R, et al. 1987. Detection of antibody to canine parvovirus in dog by enzyme immunoassay. Australian Vet J, 64: 220–221.
Teramoto Y A, Mildbrand M M, Carlson J, et al. 1984. Comparison of enzyme-linked immunosorbent assay, DNA hybridization, hemagglutination and electron microscopy for detection of canine parvovirus infections. J Clin Microbiol, 20(3): 373–378.
Waner T, Naveh A, Wudovsky I, et al. 1996. Assessment of maternal antibody decay and response to canine parvovirus vaccination using a clinic-based enzyme linked immunosorbent assay. J Vet Diagn Invest, 8: 427–432.
Waner T, Mazar S, Keren-Kornblatt E. 2006. Application of a dot ELISA for evaluation of the immune status to canine parvovirus and distemper virus in adult dog before vaccination. J Vet Diagn Invest, 18: 267–270.
Rights and permissions
About this article
Cite this article
Kumar, M., Nandi, S. & Chidri, S. Development of a polyclonal antibody-based AC-ELISA and its comparison with PCR for diagnosis of canine parvovirus infection. Virol. Sin. 25, 352–360 (2010). https://doi.org/10.1007/s12250-010-3132-x
- Canine parvovirus (CPV)
- Polyclonal antibody
- Antigen-capture ELISA(AC-ELISA)