Malik RAIES UL ISLAM, Mohinder PRATAP GUPTA, Pritam KAUR SIDHU, Gursimran FILIA, Hari MOHAN SAXENA, Tawheed AHMED SHAFI

Comparative evaluation of indirect enzyme linked immunosorbent assay, rose bengal plate test, microagglutination test, and polymerase chain reaction for diagnosis of brucellosis in buffaloes

In the present study, 178 blood samples from buffaloes were tested against indirect enzyme linked immunosorbent assay (I-ELISA), rose bengal plate test (RBPT), microagglutination test (MAT), modified microagglutination test (mMAT), and polymerase chain reaction (PCR) to select the most suitable test for efficient and effective diagnosis of bovine brucellosis. Win Episcope 2 software was used to determine the agreement between tests (kappa values at 95% confidence interval). I-ELISA was pair compared with all the other tests. Out of 178 samples, 102 were found positive by I-ELISA, 81 by RBPT, 85 by MAT, 79 by mMAT, and 68 by PCR. Substantial agreement was observed between I-ELISA and RBPT (k = 0.72), I-ELISA and MAT ((k = 0.65), and I-ELISA and mMAT ((k = 0.67). The least degree of agreement was observed between I-ELISA and PCR ((k = 0.15). I-ELISA detected more samples as positive among these tests. The results of the present study indicate that I-ELISA can be used for routine sero-diagnosis of Brucella infection in buffaloes. Furthermore, PCR can be used in combination with I-ELISA to complement the serological diagnosis, especially in the initial phase when the immune response of the animal is not detectable.

Anahtar Kelimeler:

Key words: Brucellosis, I-ELISA, RBPT, MAT, mMAT, PCR

Comparative evaluation of indirect enzyme linked immunosorbent assay, rose bengal plate test, microagglutination test, and polymerase chain reaction for diagnosis of brucellosis in buffaloes

Keywords:

I-ELISA, RBPT, MAT, mMAT, PCR, Brucellosis,

PDF

___

Garrity, G.M.: Bergey’s Manual of Systematic Bacteriology, 2nd edn. Springer, New York. 2001.
Osterman, B., Moriyon, I.: International Committee on Systematics of Prokaryotes Subcommittee on the taxonomy of Brucella. International Journal of Systematic and Evolutionary Microbiology, 2006; 56: 1173–1175.
Foster, G., Osterman, B.S., Godfroid, J., Jacques, I., Cloeckaert, A.: Brucella ceti sp. nov. and Brucella pinnipedialis sp. nov. for Brucella strains with cetaceans and seals as their preferred hosts. International Journal of Systematic and Evolutionary Microbiology, 2007; 57: 2688–93.
Enright, F.M., Walker, J.V., Jeffers, G., Deyoe, B.L.: Cellular and humoral responses of Brucella abortus-infected bovine fetuses. Am. J. Vet. Res., 1984; 45: 424–443.
Leal-Klevezas, D.S., Martinez, V.I.O., Lopez, M.A., Martinez, S.J.P.: Single step PCR for detection of Brucella spp. from blood and milk of infected animals. J. Clin. Microbiol., 1995; 3: 3087– 30
Bricker, B.J.: PCR as a diagnostic tool for brucellosis. Vet. Microbiol., 2002; 90: 435–446.
Nielsen, K., Heck, F., Wagner, G., Stiller, J., Rosenbaum, B., Pugh, R., Flores, E.: Comparative assessment of antibody isotypes to Brucella abortus by primary and secondary binding assays. Prev. Vet. Med., 1984; 2: 197–204.
Nielsen, K.: Diagnosis of brucellosis by serology. Vet. Microbiol., 2002; 90: 447–459.
Alton, G., Jones, L.M., Angus, R.D., Verger, J.M.: Techniques for the Brucellosis Laboratory. INRA, Paris. 1988.
Nasir, A.A., Parveen, Z., Ikram Ul Haq, M.: Comparative study of standard and modified serum agglutination tests for the diagnosis of brucellosis in animals. Pak. Vet. Journal, 2005; 1(25): 33–34.
OIE: Manual of the Diagnostic Tests and Vaccines for Terrestrial Animals, vol. 1, 5th edn. Office International Des Epizooties, Paris, France. 2008.
Thrusfield, M.: Veterinary Epidemiology. 3rd edn. Blackwell Science Ltd., Oxford, UK. 2005.
Corbel, M.J.: Characterization of antibodies active in Rose Bengal Plate test for bovine brucellosis. Vet. Rec., 1972; 90: 484–485.
Corbel, M. J.: Comparison of Brucella abortus and B. melitensis antigens for the Rose Bengal plate test on sera from cattle infected with B. abortus biovar-5. Vet. Rec., 1985; 117(15): 385–386.
OIE: Bovine brucellosis, Section 2.3. In OIE Manual of Standards for Diagnostic Tests and Vaccines, 5th edn. OIE, Paris. 2004.
Chachra, D., Saxena, H.M., Kaur, G., Chandra, M.: Comparative efficacy of Rose Bengal plate test, standard tube agglutination test and Dot ELISA in immunological detection of antibodies to Brucella abortus in sera. J. Bacteriology Research, 2009; 1(3): 30–
Ruppanne, R., Meyer, M.E., Willeberg, P., Behymer, D.E.: Comparison of the enzyme linked immunosorbent assay with other tests for brucellosis, using sera from experimentally infected heifers. Am. J. Vet. Research, 1980; 41: 8.
Kerby, P.J., Quiroga, J.L., McGrane, J.J., Stagg, D.A.: Field evaluation of an indirect ELISA for detection of brucellosis in lowland Bolivia. Tropical Animal Health and Production, 1997; 29: 65–72.
Rao, T.S., Devi, V.R., Babu, R.M., Rao, A.V.N.: Comparison of rapid plate agglutination, standard tube agglutination and dot-ELISA tests for the detection of antibodies to Brucella in bovines. Indian Vet. Journal, 1999; 76: 255–56.
Paweska, J.T., Potts, A.D., Harris, H.J., Smith, S.J., Viljoen., G.J., Dungu, B., Brett, O.L., Bubb, M., Prozesky, L.: Validation of an indirect enzyme-linked immunosorbent assay for the detection of antibody against Brucella abortus in cattle sera using an automated ELISA workstation. Onderstepoort J. Vet. Research, 2002; 69: 61–77.
Mittal, V., Kumar, M., Ambwani, T.: Seroepidemiological pattern of brucellosis among livestock of district Udham Singh Nagar in Uttaranchal. Indian J. Vet. Med., 2005; 25: 28–32.
Erdenebaatar, J., Bayarsaikhan, B., Yondondorj, A., Watarai, M., Shirahata, T., Jargalsaikhan, E., Kawamoto, K., Makino, S.: Epidemiological and serological survey of Brucellosis in Mongolia by ELISA using sarcosine extracts. Microbiology Immunology, 2004; 48: 571–77.
Chand, P., Sharma, A.K.: Situation of brucellosis in bovines at organized cattle farms belonging to three different states. Journal of Immunology and Immunopathology, 2004; 6: 11–
Johnson, C.A., Walker, R.D.: Clinical signs and diagnosis of Brucella canis infection. Compendium on Continuing Education for the Practicing Veterinarian, 1992; 14: 763–772.