Development and Standardization of Dot - ELISA for Detection of Neospora caninum Antibodies in Cattle and Comparison with Standard Indirect ELISA and Direct Agglutination Test (DAT).
AbstractBackground:Neospora caninum is a protozoan parasite from phylum apicomplexa and an important agent causing abortion in cattle which produce notable economic loss all around the world.Methods:Dot-Elisa was set up performing checker board procedure and then 178 sera of cattle examined with commercial indirect ELISA and direct agglutination test (DAT) were also evaluated by dot-ELISA afterwards. Results:Kappa statistical analysis revealed that Dot-ELISA has good agreements with ELISA as well as the DAT and also, Mc Nemar´s analyzing showed that this procedure has acceptable ability to discriminate positive results. Relative sensitivity and specificity of Dot-ELISA were respectively 92.63% and 89.16% and 93.4% and 90.8% in comparison with ELISA and DAT. Conclusion:Since the dot-ELISA is easy, inexpensive and not needed high experience to interpret the results, it is superior to ELISA and DAT when we aim to screen the cattle on the farm and slaughterhouses or when the laboratory equipment is not available.
Dubey JP, Schares G. Diagnosis of bovine neosporosis. Vet Parasitol. 2006; 140: 1–34.
Dubey JP, Schares G. Neosporosis in animals-the last five years. Vet Parasitol. 2011; 4(1-2): 90-108.
Anderson M, Barr B, Rowe J, Conrad P. Ne-osporosis in dairy cattle. Japn J Vet Res. 2012; 60: 51-4.
Bjerkas I, Mohn SF, Presthus J. Unidentified cyst-forming sporozoon causing encephalomy-elitis and myositis in dogs. Z Parasitenk. 1984; 70: 271–274.
McAllister MM, Dubey JP, Lindsay DS, Jolley WR, Wills RA, McGuire AM. Dogs are defini-tive hosts of Neospora caninum. Int J Parasitol. 1998; 28: 1473–1478.
Gondim LFP, McAllister MM, Pitt WC, Zem-licka DE. Coyotes (Canis latrans) are definitive hosts of Neospora caninum. Int J Parasitol. 2004; 34: 159–161.
Klevar S. Tissue cyst forming coccidia; Toxo-plasma gondii and Neospora caninum as a cause of disease in farm animals. Acta Vet Scand. 2007; 49: (Suppl1): S1.
Dubey JP. Recent advances in Neospora and neosporosis. Vet Parasitol. 1999; 84: 349-367.
Hall CA, Reichel MP, Ellis JT. Neospora abor-tions in dairy cattle: diagnosis, mode of trans-mission and control. Vet Parasitol. 2005; 128: 231–241.
Trees AJ, Williams DJL. Endogenous and ex-ogenous transplacental infection in Neospora caninum and Toxoplasma gondii. Trends Parasitol. 2005; 21: 558-561.
Björkman C, Uggla A. Serological diagnosis of Neospora caninum infection. Int J Parasitol. 1999; 29: 1497-1507.
Hamidinejat H, Ghorbanpour M, Nabavi L, Haji Hajikolaie MR, Razi Jalali MH. Occur-rence of anti-Toxoplasma gondii antibodies in fe-male cattle in south-west of Iran. Trop Anim Health Prod. 2010; 42: 899–903.
Romand S, Thulliez P, Dubey JP. Direct agglu-tination test for serologic diagnosis of Neospora caninum infection. Parasitol Res. 1998; 84: 50-3.
Desmonts G, Remington JS. Direct agglutina-tion test for diagnosis of Toxoplasma infection: method for increasing sensitivity and specific-ity. J Clin Microbiol. 1980; 11: 562–568.
Dubey JP, Desmonts G. Serological responses of equids fed Toxoplasma gondii oocysts. Equine Vet J. 1987; 19: 337-339.
Gilot-Fromont E, Aubert D, Belkilani S, Her-mitte P, Gibout O, Geers R, Villena I. Land-scape, herd management and within-herd sero-prevalence of Toxoplasma gondii in beef cattle herds from Champagne-Ardenne, France.Vet Parasitol. 2009; 161: 36–40.
Dohoo IR, Martin SW, Stryhn H. In: Dohoo IR. (Ed.). Vet Epidemiol Res. AVC Inc., Char-lottetown, 2003; p: 706.
Kemeny DM. Titration of antibodies. J Immu-nol Methods. 1992; 150: 57-76.
Wouda W, Brinkhof J, Van Maanen C, de Gee Al, Moen AR. Serodiagnosis of neosporosis in individual cows and dairy herds: a comparative study of three enzyme-linked immunosorbent assays. Clin Diagn Lab Immunol. 1998; 5: 711–716.
dos Santos DS, Andrade MP, Varaschin MS, Guimarães AM, Hirsch C. Neospora caninum in bovine fetuses of Minas Gerais, Brazil: genetic characteristics of rDNA. Rev Bras Parasitol Vet. 2011; 20: 281-8.
Leon A, Richard E, Fortier C, Laugier C, Forti-er G, Pronost S. Molecular detection of Coxiella burnetii and Neospora caninum in equine aborted foetuses and neonates. Prev Vet Med. 2012; 104(1-2): 179-83.
Jafar Pour Azami S, Keshavarz H, Rezaian M, Mohebali M, Shojaee S. Rapid Detection of Toxoplasma gondii Antigen in Experimentally In-fected Mice by Dot- ELISA. Iranian J Parasitol. 2011; 6: 28-33.
Huang P, Liao M, Zhang H, Lee EG, Nishi-kawa Y, Xuan X. Dense-granule protein Nc GRA7, a new marker for the serodiagnosis of Neospora caninum infection in aborting cows. Clin Vaccine Immunol. 2007; 14: 1640-3.
Kwok OC, Dubey JP. Evaluation of four sero-logical techniques to determine the seroprev-alence of Neospora caninum in foxes (Vulpes vul-pes) and coyotes (Canis latrans) on Prince Ed-ward Island, Canada. Vet Parasitol. 2007; 10: 51-8.
Wapenaar W, Barkema HW, Vanleeuwen JA, McClure JT, O'Handley RM, Kwok OC, Thulliez P, Dubey JP, Jenkins MC. Compari-son of serological methods for the diagnosis of Neospora caninum infection in cattle. Vet Parasi-tol. 2007; 143(2):166-73.
Aguado-Martínez A, Alvarez-García G, Fer-nández-García A, Risco-Castillo V, Arnaiz-Seco I, Rebordosa-Trigueros X, Navarro-Lozano V, Ortega-Mora LM. Usefulness of rNcGRA7- and rNcSAG4-based ELISA tests for distinguishing primo-infection, recrudes-cence, and chronic bovine neosporosis. Vet Parasitol. 2008; 157: 182-95.
Hu J, Ferroglio E, Trisciuoglio A. Immunoblot diagnosis of infection with Neospora caninum in cattle based on recombinant NcSAG4 Antigen. Parasitol Res. 2011; 108: 1055-8.
Nematollahi A, Jaafari Jozani R, Zaboli N. Ad-aptation of Dot-Elisa for Serodiagnosis of Ne-ospora caninum Infestation in Aborted Cows. Glob Veterinaria. 2011; 7 (2): 149-152.
Dangoudoubiyam S, Oliveira JB, Víquez C, Gómez-García A, González O, Romero JJ, Kwok OC, Dubey JP, Howe DK. Detection of antibodies against Sarcocystis neurona, Neospora spp., and Toxoplasma gondii in horses from Cos-ta Rica. J Parasitol. 2011; 97: 522-4.
Packham AE, Sverlow KW, Conrad PA, Loomis EF, Rowe JD, Anderson ML, Marsh AE, Cray C. Barr BC. A modified agglutination test for Neospora caninum: development, optimi-zation, and comparison to the indirect fluo-rescent-antibody test and enzyme-linked im-munosorbent assay. Clin Diagn Lab Immunol. 1998; 5: 467–473.
Pinheiro AM, Costa MF, Paule B, Vale V, Ri-beiro M, Nascimento I, Schaer RE, Almeida MA, Meyer R, Freire SM. Serologic immuno-reactivity to Neospora caninum antigens in dogs determined by indirect immunofluorescence, western blotting and dot-ELISA. Vet Parasitol. 2005; 130 : 73–79.
Salehi N, Haddadzadeh HR, Shayan P, Vodjgani M, Bolourchi M. Serological study of Neospora caninum in pregnant dairy cattle in Teh-ran, Iran. Int J Vet Res. 2010; 4: 113-116.
Haji Hajikolaei MR, Hamidinejat H, Ghor-banpoor M, Goraninejad S. Serological study of Neospora caninum infection in cattle from Ah-vaz area, Iran. Int J Vet Res. 2008; 2: 63-66.
Nematollahi A, Jaafari R, Moghaddam G. Se-roprevalence of Neospora caninum Infection in Dairy Cattle in Tabriz, Northwest Iran. Iranian J Parasitol. 2011; 6: 95-8.
Razmi GR, Mohammadi GR, Garrosi T, Far-zaneh N, Fallah AH, Maleki M. Seroepidemiol-ogy of Neospora caninum infection in dairy cattle herds in Mashhad area, Iran. Vet Parasitol. 2006; 30: 187-9.
Nourollahi Fard SR, Khalili M, Aminzadeh A. Prevalence of antibodies to Neospora caninum in cattle in Kerman province, South East Iran.Veterinarski Arhiv. 2008; 78: 253-259.
Sharifdini M, Mohebali M, Keshavarz H, Hos-seininejad M, Hajjaran H, Akhoundi B, Rahimi Foroushani A, Zarei Z, Charehdar S. Neospora caninum and Leishmania infantum co-infection in domestic dogs (Canis familiaris) in Meshkin-Shahr district, Northwestern Iran. Iran J Arthropod-Borne Dis. 2011; 1(3): 60–68.