Comparative study of in vitro prepared Rose Bengal Plate Test (RBPT) antigen with commercially available antigens

Mujeeb ur Rahman 1 * , Amir Ullah 2, Junaid Ali Shah 3
More Detail
1 College of Life Sciences, Northwest University, Xi’an, Shaanxi province, China
2 Department of Laboratory Medicine, Southern Medical University, Guangzhou, China
3 College of Life Sciences, Jilin University, China Changchun city, Jilin Province
* Corresponding Author
J CLIN MED KAZ, Volume 5, Issue 59, pp. 46-50.
Download Full Text (PDF)


Background and aim: Brucellosis is one of the world most common zoonotic diseases. The current study was aimed to prepare the Rose Bengal Plate Test (RBPT) antigen for the diagnosis of brucellosis and to determine its specificity and sensitivity.
Material and methods: The Rose Bengal plate test antigen prepared from Brucella abortus (B. abortus) strain 99 was compared with two commercial Rose Bengal Plate Test antigens and its specificity and sensitivity are determined.
Results: The were Rose Bengal plate test and I-ELISA result show that the in vitro antigen was superior to RBPT antigen University Diagnosis Laboratory (UDL) Lahore Pakistan, and RBPT antigen Veterinary Laboratory Agency (VLA) UK. Out of 196 samples analyzed by in vitro RBPT antigen, RBPT antigen (UDL), RBPT antigen (VLA), and an indirect enzyme-linked immunosorbent assay (I-ELISA) 56.63 %, 53.57%, 41.84%, 35.71% were found B. abortus positively. The sensitivity calculated for the in vitro RBPT antigen was 96.62, while RBPT antigen (UDL) and RBPT antigen (VLA) were 89.77, 63.91 correspondingly. However, the specificity of the in vitro RBPT antigen was lower (77.57%), than the commercial RBPT antigen (VLA) (79.79%).
Conclusions: A very sensitive and low-cost in vitro RBPT antigen compared to commercial RBPT was magnificently developed in the current study. It was determined that the in vitro RBPT antigen could substitute the available commercial RBPT antigen, which is comparatively expensive and less sensitive in the detection of brucellosis. Therefore, it is concluded that the in vitro RBPT antigen could be used for epidemiological surveillance of brucellosis.


Rahman MU, Ullah A, Shah JA. Comparative study of in vitro prepared Rose Bengal Plate Test (RBPT) antigen with commercially available antigens. J Clin Med Kaz. 2020;5(59):46-50.


  • Godfroid J, Scholz H, Barbier T, Nicolas C, Wattiau P, Fretin D, et al. Brucellosis at the animal/ecosystem/human interface at the beginning of the 21st century. Preventive veterinary medicine. 2011; 102(2):118-131.
  • Scholz HC, Revilla-Fernández S, Al Dahouk S, Hammerl JA, Zygmunt MS, Cloeckaert A, et al. Brucella vulpis sp. nov., isolated from mandibular lymph nodes of red foxes (Vulpes vulpes). International journal of systematic and evolutionary microbiology. 2016; 66(5):2090-2098.
  • Aparicio ED. Epidemiology of brucellosis in domestic animals caused by Brucella melitensis, Brucella suis and Brucella abortus. Rev Sci Tech. 2013; 32(1):53-60.
  • Saleem MZ, Akhtar R, Aslam A, Rashid MI, Chaudhry ZI, Manzoor MA, et al. Evidence of Brucella abortus in non-preferred caprine and ovine hosts by real-time PCR assay. Pak J Zool. 2019; 51:1187-1189.
  • Saeed U, Ali S, Khan TM, El-Adawy H, Melzer F, Khan AU, et al. Seroepidemiology and the molecular detection of animal brucellosis in Punjab, Pakistan. Microorganisms. 2019; 7(10):449.
  • Jamil T, Melzer F, Khan I, Iqbal M, Saqib M, Hammad Hussain M, et al. Serological and molecular investigation of Brucella species in dogs in Pakistan. Pathogens. 2019; 8(4):294.
  • Fatima S, Khan I, Nasir A, Younus M, Saqib M, Melzer F, et al. Serological, molecular detection and potential risk factors associated with camel brucellosis in Pakistan. Tropical animal health and production. 2016; 48(8):1711-1718.
  • Foster JT, Walker FM, Rannals BD, Hussain MH, Drees KP, Tiller RV, et al. African lineage Brucella melitensis isolates from Omani livestock. Frontiers in microbiology. 2018; 8:2702.
  • Norman FF, Monge-Maillo B, Chamorro-Tojeiro S, Pérez-Molina J-A, López-Vélez R. Imported brucellosis: a case series and literature review. Travel medicine and infectious disease. 2016; 14(3):182-199.
  • Neha A, Kumar A, Ahmed I. Comparative efficacy of serological diagnostic methods and evaluation of polymerase chain reaction for diagnosis of bovine brucellosis. Iranian journal of veterinary research. 2017; 18(4):279.
  • Franc K, Krecek R, Häsler B, Arenas-Gamboa A. Brucellosis remains a neglected disease in the developing world: a call for interdisciplinary action. BMC public health. 2018; 18(1):1-9.
  • Seleem MN, Boyle SM, Sriranganathan N. Brucellosis: a re-emerging zoonosis. Veterinary microbiology. 2010; 140(3-4):392-398.
  • Corbel MJ. Brucellosis in humans and animals: World Health Organization; 2006.
  • Akhtar S, Mirza M. Rates of seroconversion in the progeny of Brucella abortus seropositive and seronegative cattle and buffalo. Revue scientifique et technique (International Office of Epizootics). 1995; 14(3):711-718.
  • Dadar M, Shahali Y, Whatmore AM. Human brucellosis caused by raw dairy products: A review on the occurrence, major risk factors and prevention. International journal of food microbiology. 2019; 292:39-47.
  • Hakeem M, Saeed S. Brucellosis: A case report and literature review. J Postgrad Med Ed Res. 2019; 53:126-127.
  • Young E, Hall W, Evans A, Brachman P. Bacterial infection of humans: epidemiology and control. 1998.
  • Rubach MP, Halliday JE, Cleaveland S, Crump JA. Brucellosis in low-income and middle-income countries. Current opinion in infectious diseases. 2013; 26(5):404.
  • Glynn MK, Lynn TV. Zoonosis update. AVMA. 2008; 233:900-908.
  • Muñoz P, Marín C, Monreal D, Gonzalez D, Garin-Bastuji B, Diaz R, et al. Efficacy of several serological tests and antigens for diagnosis of bovine brucellosis in the presence of false-positive serological results due to Yersinia enterocolitica O: 9. Clinical and Diagnostic Laboratory Immunology. 2005; 12(1):141-151.
  • Refai M. Incidence and control of brucellosis in the Near East region. Veterinary microbiology. 2002; 90(1-4):81-110.
  • Padilla PF, Nielsen K, Ernesto SL, Ling YW. Diagnosis of brucellosis. The Open Veterinary Science Journal. 2010; 4(1):46-60.
  • Morgan WB, MacKinnon D, Cullen G. Rose Bengal plate agglutination test in the diagnosis of brucellosis. Veterinary record. 1969.
  • Oomen L, Waghela S. The rose bengal plate test in human brucellosis. Tropical and geographical medicine. 1974; 26(3):300-302.
  • Alexander D. Newcastle, disease: OIE Terrestrial Manual 2009. Manual of diagnostic tests and vaccines for terrestrial animals. 2009:3.
  • Akhtar R, Chaudhry ZI, Shakoori A, Ahmad M, Aslam A. Comparative efficacy of conventional diagnostic methods and evaluation of polymerase chain reaction for the diagnosis of bovine brucellosis. Vet World. 2010; 3(2):53-56.
  • Abbas BA, Mohammed HQ, Tofah JA. Diagnostic Study on Brucella melitensis isolated from Human in Wassit province. Al-Mustansiriyah Journal for Pharmaceutical Sciences. 2012; 12(2):8-24.
  • Gilligan P, York M, Baselski V, Carey R, Krisher K, Lovchik J, et al. Sentinel Laboratory Guidelines for Suspected Agents of Bioterrorism: Brucella Species. Washington, ASM Press. 2004; 1:4-10.
  • Noviello S, Gallo R, Kelly M, Limberger RJ, DeAngelis K, Cain L, et al. Laboratory-acquired brucellosis. Emerging infectious diseases. 2004; 10(10):1848.
  • Zakaria AM. Comparative Assessment of Sensitivity and Specificity of Rose Bengal Test and Modified In-House ELISA by using IS711 Taqman Real Time PCR Assay as a Gold Standard for the Diagnosis of Bovine Brucellosis. Biomedical and Pharmacology Journal. 2018; 11(2):951-957.
  • Shahaza O, Khairani-Bejo S, Zunita Z, Bahaman A. In-house Rose Bengal Plate Agglutination Test (RBPT) for a rapid diagnosis of brucellosis in goats in Malaysia. Int J Trop Med. 2009; 4(3):116-118.
  • Saxena HM, Chothe S, Kaur P. Simple solutions to false results with plate/slide agglutination tests in diagnosis of infectious diseases of man and animals. MethodsX. 2015; 2:345-352.
  • Manishimwe R, Ntaganda J, Habimana R, Nishimwe K, Byukusenge M, Dutuze F, et al. Comparison between Rose Bengal Plate Test and competitive enzyme linked immunosorbent assay to detect bovine brucellosis in Kigali City, Rwanda. J Vet Sci Technol. 2015; 6:1-4.
  • Yahaya SM, Khairani-Bejo S, Zunita Z, Omar MA, Bitrus AA. Development of an in-house Rose Bengal Plate test for diagnosis of brucellosis in goat. Malaysian Journal of Veterinary Research. 2016; 7(2):59-65.