Development of latex agglutination test with recombinant NcSAG1 for the rapid detection of antibodies to Neospora caninum in cattle.

Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. Surface antigen 1 of N. caninum (NcSAG1) is an important immunodominant candidate for the development of a diagnostic reagent for neosporosis. The present study describes the development and evaluation of a latex agglutination test (LAT) with recombinant NcSAG1 (rNcSAG1) for the detection of antibodies to N. caninum in cattle. The rNcSAG1 gene was cloned in pET-28a and protein was expressed in Escherichia coli BL21 (DE3). Carboxylated latex particles were coated with rNcSAG1 and the degree of agreement between LAT and a commercial enzyme-linked immunosorbent assay (iscomELISA) was evaluated by using of 164 serum samples. Twenty-two (13.4%) and 23 (14.0%) of samples were positive for antibodies to N. caninum by LAT and ELISA respectively. Eighteen of 23 ELISA-positive samples were positive according to the LAT and a substantial agreement (κ=0.77) was found between the results of LAT and ELISA. The results indicated that the LAT with rNcSAG1 would be a rapid, simple, relatively inexpensive and suitable diagnostic test for detection of specific antibodies in N. caninum infection under field conditions. Improvement in purification of rNcSAG1 can reduce probable false positive reactions and so increase the degree of agreement between the LAT and ELISA.

[1]  Xiaohong Huang,et al.  Development of Rapid Immunochromatographic Test with Recombinant NcSAG1 for Detection of Antibodies to Neospora caninum in Cattle , 2005, Clinical Diagnostic Laboratory Immunology.

[2]  A. Hemphill,et al.  Identification and partial characterization of a 36 kDa surface protein on Neospora caninum tachyzoites , 1997, Parasitology.

[3]  M. Reichel,et al.  Progress in the serodiagnosis of Neospora caninum infections of cattle. , 2000, Parasitology today.

[4]  G. Hill,et al.  Latex Agglutination Reactions Between Human Chorionic Gonadotropin and Rabbit Antibody.∗ , 1962, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[5]  L. Sibley,et al.  The p29 and p35 Immunodominant Antigens ofNeospora caninum Tachyzoites Are Homologous to the Family of Surface Antigens of Toxoplasma gondii , 1998, Infection and Immunity.

[6]  D. Wiedbrauk,et al.  Latex agglutination test for detection of antibodies to Toxoplasma gondii , 1988, Journal of clinical microbiology.

[7]  C. Plotz,et al.  The latex fixation test. I. Application to the serologic diagnosis of rheumatoid arthritis. , 1956, The American journal of medicine.

[8]  S. Matsuzawa,et al.  Microtiter latex antiglobulin test for the detection of antibodies to DNP, digoxin, HCG and similar antigens. , 1983, Journal of immunological methods.

[9]  P. Lowe,et al.  Purification of Calf Prochymosin (Prorennin) Synthesized in Escherichia Coli , 1984, Bio/Technology.

[10]  P. Conrad,et al.  Cloning and characterization of two recombinant Neospora protein fragments and their use in serodiagnosis of bovine neosporosis , 1997, Clinical and diagnostic laboratory immunology.

[11]  Á. Zaballos,et al.  The NcGRA7 gene encodes the immunodominant 17 kDa antigen of Neospora caninum , 2006, Parasitology.

[12]  Hiroshi Suzuki,et al.  Serodiagnosis of Neospora caninum infection in cattle by enzyme-linked immunosorbent assay with recombinant truncated NcSAG1. , 2003, Veterinary parasitology.

[13]  T. Jiang,et al.  Evaluation of a recombinant MIC3 based latex agglutination test for the rapid serodiagnosis of Toxoplasma gondii infection in swines. , 2008, Veterinary parasitology.

[14]  G. Schares,et al.  Epidemiology and Control of Neosporosis and Neospora caninum , 2007, Clinical Microbiology Reviews.

[15]  D. Buxton,et al.  An IgG Avidity ELISA to Discriminate between Recent and Chronic Neospora Caninum Infection , 1999, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[16]  M. Puterman,et al.  Evaluation of seven immunoassays for detection of rotavirus in pediatric stool samples , 1988, Journal of clinical microbiology.

[17]  M. Thurmond,et al.  Detection of Serum Antibody Responses in Cattle with Natural or Experimental Neospora Infections , 1993, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[18]  J. Dubey,et al.  Evaluation of two Neospora caninum recombinant antigens for use in an enzyme-linked immunosorbent assay for the diagnosis of bovine neosporosis , 1996, Clinical and diagnostic laboratory immunology.

[19]  Diana J L Williams,et al.  Diagnosis and seroepidemiology of Neospora caninum-associated bovine abortion. , 2002, International journal for parasitology.

[20]  C. Björkman,et al.  Serological diagnosis of Neospora caninum infection. , 1999, International journal for parasitology.

[21]  C. Sugimoto,et al.  Serodiagnosis of Toxoplasma gondii infection in cats by enzyme-linked immunosorbent assay using recombinant SAG1. , 2001, Veterinary parasitology.

[22]  J. Dubey,et al.  Review of Neospora caninum and neosporosis in animals. , 2003, The Korean journal of parasitology.

[23]  D. Lindsay,et al.  Neonatal Neospora caninum infection in dogs: isolation of the causative agent and experimental transmission. , 1988, Journal of the American Veterinary Medical Association.

[24]  R. Z. Machado,et al.  Neospora caninum infection in birds: experimental infections in chicken and embryonated eggs , 2007, Parasitology.

[25]  D. Lindsay,et al.  A review of Neospora caninum and neosporosis. , 1996, Veterinary parasitology.

[26]  J. Dubey,et al.  Diagnosis of bovine neosporosis. , 2006, Veterinary parasitology.

[27]  P. Conrad,et al.  Evidence of vertical transmission of Neospora sp infection in dairy cattle. , 1997, Journal of the American Veterinary Medical Association.

[28]  A. Niveleau,et al.  The preparation of latex particles with covalently bound polyamines, IgG and measles agglutinins and their use in visual agglutination tests. , 1978, Journal of immunological methods.

[29]  L. F. Gondim,et al.  Coyotes (Canis latrans) are definitive hosts of Neospora caninum. , 2004, International journal for parasitology.

[30]  Xiaohong Huang,et al.  Rapid Immunochromatographic Test Using Recombinant SAG2 for Detection of Antibodies against Toxoplasma gondii in Cats , 2004, Journal of Clinical Microbiology.

[31]  F. Gella,et al.  Latex agglutination procedures in immunodiagnosis , 1991 .

[32]  J. Mineo,et al.  Evaluation of serological tests for the diagnosis of Neospora caninum infection in dogs: optimization of cut off titers and inhibition studies of cross-reactivity with Toxoplasma gondii. , 2007, Veterinary parasitology.

[33]  D. Lindsay,et al.  Dogs are definitive hosts of Neospora caninum. , 1998, International journal for parasitology.

[34]  D. Moore,et al.  An applied printing immunoassay with recombinant Nc-SAG1 for detection of antibodies to Neospora caninum in cattle , 2011, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[35]  P. Conrad,et al.  A Modified Agglutination Test for Neospora caninum: Development, Optimization, and Comparison to the Indirect Fluorescent-Antibody Test and Enzyme-Linked Immunosorbent Assay , 1998, Clinical Diagnostic Laboratory Immunology.

[36]  J. Dubey,et al.  Serological diagnosis of bovine neosporosis by Neospora caninum monoclonal antibody-based competitive inhibition enzyme-linked immunosorbent assay , 1996, Journal of clinical microbiology.

[37]  Y. Sukthana,et al.  Predictive value of latex agglutination test in serological screening for Toxoplasma gondii. , 2001, The Southeast Asian journal of tropical medicine and public health.

[38]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[39]  J. Wastling,et al.  A multiple antigen ELISA to detect Neospora-specific antibodies in bovine sera, bovine foetal fluids, ovine and caprine sera. , 1998, Veterinary parasitology.