Hostname: page-component-78c5997874-4rdpn Total loading time: 0 Render date: 2024-11-05T16:08:34.789Z Has data issue: false hasContentIssue false

Development of a practical immunochromatographic test with recombinant P50 for the diagnosis of Babesia gibsoni infection in dogs

Published online by Cambridge University Press:  22 July 2005

R. A. VERDIDA
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
X. XUAN
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
S. FUKUMOTO
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
X. HUANG
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
J. ZHOU
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
I. IGARASHI
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
F. G. CLAVERIA
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
H. NAGASAWA
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan

Abstract

An immunochromatographic test (ICT), using recombinant truncated P50 (P50t), for the detection of antibodies to Babesia gibsoni was developed and evaluated. Whereas all sera from specific pathogen-free dogs were clearly negative, all sera from dogs experimentally infected with B. gibsoni were clearly positive in the ICT. In addition, the ICT detected no cross-reactivity with sera from dogs experimentally infected with closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi, or with Neospora caninum, and Leishmania infantum. Sequential sera from a dog experimentally infected with B. gibsoni were tested with the ICT; it was shown that the specific antibodies are detectable as early as 6 days post-infection (p.i.) and that strong antibody responses remained until the end of the experiment (144 days p.i.). To evaluate the clinical application of the ICT, a total of 54 serum samples collected from domestic dogs that had been identified as having signs of anaemia at veterinary hospitals in Japan, were tested with the ICT, the previously established enzyme-linked immunosorbent assay (ELISA) and with the indirect fluorescent antibody test (IFAT). Twenty-four of the tested samples (44·4%) were positive in both ICT and ELISA, and (51·8%) in IFAT. The concordance between ELISA and ICT was found to be 100%, and 85·7% with IFAT. Taken together, the results above suggest that the ICT using P50t is rapid, simple, accurate, and suitable for use at clinical sites for the diagnosis of B. gibsoni infection in dogs.

Type
Research Article
Copyright
© 2005 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Birkenheuer, A. J., Levy, M. G. and Breitschwerdt, E. ( 2003 a). Development and evaluation of a seminested PCR for detection and differentiation of Babesia gibsoni (Asian genotype) and B. canis DNA in canine blood samples. Journal of Clinical Microbiology 41, 41724177.Google Scholar
Birkenheuer, A. J., Levy, M. G., Stebbins, M., Poore, M. and Breitschwerdt, E. ( 2003 b). Serosurvey of anti- Babesia antibodies in stray dogs and American Pit Bull Terriers and American Staffordshire Terriers from North Carolina. Journal of the American Animal Hospital Association 39, 551557.Google Scholar
Boozer, A. L. and Macintire, D. K. ( 2003). Canine babesiosis. Veterinary Clinics Small Animal Practice 33, 885904.CrossRefGoogle Scholar
Bose, R., Jorgensen, W. K., Dalgliesh, R. J., Friedhoff, K. T. and De Vos, A. J. ( 1995). Current state and future trends in the diagnosis of babesiosis. Veterinary Parasitology 57, 6174.CrossRefGoogle Scholar
Bruning, A. ( 1996). Equine piroplasmosis: an update on diagnosis, treatment, and prevention. The British Veterinary Journal 152, 139151.CrossRefGoogle Scholar
Caccio, S. M., Antunovic, B., Moretti, A., Mangili, V., Marinculic, A., Baric, R. F., Slemenda, S. B. and Pieniazek, N. J. ( 2002). Molecular characterization of Babesia canis canis and Babesia canis vogelli from naturally infected European dogs. Veterinary Parasitology 106, 285292.CrossRefGoogle Scholar
Camacho, A. T., Pallas, E., Gestal, J. J., Guitian, F. J., Olmeda, H. K., Goethert, K. and Telford, S. R. ( 2001). Infection of dogs in north-west Spain with Babesia microti-like agent. Veterinary Record 149, 552555.CrossRefGoogle Scholar
Chan, R., Chen, J., York, M. K., Setijono, N., Kaplan, R. L., Graham, F. and Tanowitz, H. B. ( 2000). Evaluation of a combination rapid immunoassay for detection of Giardia and Cryptosporidium antigens. Journal of Clinical Microbiology 38, 393394.Google Scholar
Chandler, J., Gurmin, T. and Robinson, N. ( 2000). The place of gold in rapid tests. IVD Technology 6, 3749.Google Scholar
De Waal, D. T. ( 2000). Global importance of piroplasmosis. Journal of Protozoology Research 10, 10627.Google Scholar
Fukumoto, S., Xuan, X., Nishikawa, Y., Inoue, N., Igarashi, I., Nagasawa, H., Fujisaki, K. and Mikami, T. ( 2001). Identification and expression of a 50-kilodalton surface antigen of Babesia gibsoni and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay. Journal of Clinical Microbiology 39, 26032609.CrossRefGoogle Scholar
Huang, X., Xuan, X., Hirata, H., Yokoyama, N., Xu, L., Suzuki, N. and Igarashi, I. ( 2004 a). Rapid immunochromatographic test using recombinant SAG2 for detection of antibodies against Toxoplasma gondii in cats. Journal of Clinical Microbiology 42, 351353.Google Scholar
Huang, X., Xuan, X., Xu, L., Zhang, S., Yokoyama, N., Suzuki, N. and Igarashi, I. ( 2004 b). Development of an immunochromatographic test with recombinant EMA-2 foe the rapid detection of antibodies against Babesia equi in horses. Journal of Clinical Microbiology 42, 359361.Google Scholar
Jefferies, R., Ryan, U. M., Muhlnickel, C. J. and Irwin, P. J. ( 2003). Two species of canine babesia in Australia: detection and characterization by PCR. Journal of Parasitology 89, 409412.CrossRefGoogle Scholar
Kjemtrup, A. M., Kocan, A. A., Whitworth, L., Meinkoth, J., Birkenheuer, A. J., Cummings, J., Boudreaux, M. K., Stockham, S. L., Irizarry-Rovira, A. and Conrad, P. A. ( 2000). There are at least three genetically distinct small piroplasms from dogs. International Journal for Parasitology 30, 15011505.CrossRefGoogle Scholar
Levine, N. D. ( 1985). Veterinary Protozoology. Iowa State University Press, Ames, Iowa.
Mills, C. D., Burgess, D. C. H., Taylor, H. J. and Kain, K. C. ( 1999). Evaluation of a rapid and inexpensive dipstick assay for the diagnosis of Plasmodium falciparum malaria. Bulletin of the World Health Organization 77, 553558.Google Scholar
Mohebali, M., Taran, M. and Zarei, Z. ( 2004). Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick rk39 test and direct agglutination. Veterinary Parasitology 121, 239245.CrossRefGoogle Scholar
Muhlnickel, C. J., Jefferies, R., Ryan, U. M. and Irwin, P. J. ( 2002). Babesia gibsoni infection in three dogs in Victoria. Australian Veterinary Journal 80, 606610.CrossRefGoogle Scholar
Reithinger, R., Quinell, R. J., Alexander, B. and Davies, C. R. ( 2002). Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick test, enzyme-linked immunosorbent assay, and PCR. Journal of Clinical Microbiology 40, 23522356.CrossRefGoogle Scholar
Richardson, D. C., Ciach, M., Zhong, K. J. Y., Crandall, I. and Kain, K. C. ( 2002). Evaluation of the Makromed dipstick assay versus PCR for diagnosis of Plasmodium falciparum malaria in returned travelers. Journal of Clinical Microbiology 40, 45284530.CrossRefGoogle Scholar
Shaw, S. E., Day, M. J. and Breitschwerdt, E. B. ( 2001). Tick-borne infectious diseases of dogs. Trends in Parasitology 17, 7480.CrossRefGoogle Scholar
Verdida, R. A., Hara, O. A., Xuan, X., Fukumoto, S., Igarashi, I., Zhang, S., Dong, J., Inokuma, H., Kabeya, H., Sato, Y., Moritomo, T., Maruyama, S., Claveria, F. and Nagasawa, H. ( 2004). Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50. Journal of Veterinary Medical Science 66, 15171521.CrossRefGoogle Scholar
Yamane, I., Conrad, P. and Gardner, I. ( 1993 a). Babesia gibsoni infections in dogs. Journal of Protozoology Research 3, 111125.Google Scholar
Yamane, I., Thomford, J. W., Gardner, I. A., Dubey, J. P., Levy, J. P. and Conrad, P. A. ( 1993 b). Evaluation of the indirect fluorescent antibody test for diagnosis of Babesia gibsoni infections in dogs. American Journal of Veterinary Research 54, 15791584.Google Scholar
Zahler, M., Rinder, H., Schein, E. and Gothe, R. ( 2000 a). Detection of a new pathogenic Babesia microti-like species in dogs. Veterinary Parasitology 89, 241248.Google Scholar
Zahler, M., Rinder, H., Zweygarth, E., Fukata, T., Maede, Y., Schein, E. and Gothe, R. ( 2000 b). Babesia gibsoni’ of dogs from North America and Asia belong to different species. Parasitology 120, 365369.Google Scholar