Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera
Abstract Background Brucellosis is an economically devastating animal disease and has public health concern. Serological methods such as Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), and Indirect-Enzyme-Linked Immunosorbent Assay (I-ELISA) have been used to detect brucellosis. Howev...
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| Format: | Article |
| Language: | English |
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BMC
2023-08-01
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| Series: | BMC Microbiology |
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| Online Access: | https://doi.org/10.1186/s12866-023-02962-2 |
| _version_ | 1797577787692810240 |
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| author | Abinet Legesse Aregitu Mekuriaw Esayas Gelaye Takele Abayneh Belayneh Getachew Wubet Weldemedhin Takele Tesgera Getaw Deresse Kenaw Birhanu |
| author_facet | Abinet Legesse Aregitu Mekuriaw Esayas Gelaye Takele Abayneh Belayneh Getachew Wubet Weldemedhin Takele Tesgera Getaw Deresse Kenaw Birhanu |
| author_sort | Abinet Legesse |
| collection | DOAJ |
| description | Abstract Background Brucellosis is an economically devastating animal disease and has public health concern. Serological methods such as Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), and Indirect-Enzyme-Linked Immunosorbent Assay (I-ELISA) have been used to detect brucellosis. However, there is limited comparative evaluation studies and lack of molecular confirmation of the causative agents in the study areas. The study was aimed to compare RBPT, I-ELISA, CFT, and confirmation using Polymerase Chain Reaction (PCR). A total of 2317 sera samples were collected from brucellosis-affected areas of Ethiopia with no vaccination history. All sera were subjected to comparative serological assays. Post-cross tabulation, sensitivity, and specificity were determined using Receiver Operating Characteristics (ROC) curve analysis software. PCR was performed on 54 seropositive samples using genus- and species-specific primers. Results Among the 2317 sera tested for comparative serological assays, 189 (8.16%) were positive for RBPT, 191 (8.24%) for I-ELISA, and 48 (2.07%) for CFT. Sensitivity to RBPT was 100% (95%) in shoats and 74% (95%) in cattle. Specificity on RBPT was 98.69% (95%), 99.28% (95%), 100% (95%) in sheep, goats, and cattle, respectively. CFT sensitivity was 4 (95%) in sheep, 9.65 (95%) goats, and 72 (95%) cattle. Specificity on CFT was 100% (95%) for sheep, goats, and cattle. A 223bp Brucella genus-specific and 156bp B. abortus species-specific detected. However, B. melitensis not detected. Conclusion In this study, I-ELISA was the most sensitive and specific test. RBPT detected all Brucellosis-infected sheep and goats; nevertheless, it showed false positive in sheep and goats and false negative in cattle. The presence of B. abortus in small and large ruminants was confirmed by PCR. This is the first report of B. abortus detection in small ruminant in Ethiopia. B.abortus detected in non-preferred hosts. The findings suggest further study on molecular epidemiology of Brucella species. |
| first_indexed | 2024-03-10T22:13:56Z |
| format | Article |
| id | doaj.art-faae5bf4ac5e4c86bd6c9cf216d8ec78 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-03-10T22:13:56Z |
| publishDate | 2023-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj.art-faae5bf4ac5e4c86bd6c9cf216d8ec782023-11-19T12:32:28ZengBMCBMC Microbiology1471-21802023-08-012311810.1186/s12866-023-02962-2Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle seraAbinet Legesse0Aregitu Mekuriaw1Esayas Gelaye2Takele Abayneh3Belayneh Getachew4Wubet Weldemedhin5Takele Tesgera6Getaw Deresse7Kenaw Birhanu8Research and Development Directorate, National Veterinary InstituteResearch and Development Directorate, National Veterinary InstituteFood and Agriculture Organization of the United Nations, Sub-Regional Office for Eastern AfricaResearch and Development Directorate, National Veterinary InstituteResearch and Development Directorate, National Veterinary InstituteResearch and Development Directorate, National Veterinary InstituteResearch and Development Directorate, National Veterinary InstituteResearch and Development Directorate, National Veterinary InstituteResearch and Development Directorate, National Veterinary InstituteAbstract Background Brucellosis is an economically devastating animal disease and has public health concern. Serological methods such as Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), and Indirect-Enzyme-Linked Immunosorbent Assay (I-ELISA) have been used to detect brucellosis. However, there is limited comparative evaluation studies and lack of molecular confirmation of the causative agents in the study areas. The study was aimed to compare RBPT, I-ELISA, CFT, and confirmation using Polymerase Chain Reaction (PCR). A total of 2317 sera samples were collected from brucellosis-affected areas of Ethiopia with no vaccination history. All sera were subjected to comparative serological assays. Post-cross tabulation, sensitivity, and specificity were determined using Receiver Operating Characteristics (ROC) curve analysis software. PCR was performed on 54 seropositive samples using genus- and species-specific primers. Results Among the 2317 sera tested for comparative serological assays, 189 (8.16%) were positive for RBPT, 191 (8.24%) for I-ELISA, and 48 (2.07%) for CFT. Sensitivity to RBPT was 100% (95%) in shoats and 74% (95%) in cattle. Specificity on RBPT was 98.69% (95%), 99.28% (95%), 100% (95%) in sheep, goats, and cattle, respectively. CFT sensitivity was 4 (95%) in sheep, 9.65 (95%) goats, and 72 (95%) cattle. Specificity on CFT was 100% (95%) for sheep, goats, and cattle. A 223bp Brucella genus-specific and 156bp B. abortus species-specific detected. However, B. melitensis not detected. Conclusion In this study, I-ELISA was the most sensitive and specific test. RBPT detected all Brucellosis-infected sheep and goats; nevertheless, it showed false positive in sheep and goats and false negative in cattle. The presence of B. abortus in small and large ruminants was confirmed by PCR. This is the first report of B. abortus detection in small ruminant in Ethiopia. B.abortus detected in non-preferred hosts. The findings suggest further study on molecular epidemiology of Brucella species.https://doi.org/10.1186/s12866-023-02962-2Brucella speciesDiagnosisCFTI-ELISARBPTPCR |
| spellingShingle | Abinet Legesse Aregitu Mekuriaw Esayas Gelaye Takele Abayneh Belayneh Getachew Wubet Weldemedhin Takele Tesgera Getaw Deresse Kenaw Birhanu Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera BMC Microbiology Brucella species Diagnosis CFT I-ELISA RBPT PCR |
| title | Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera |
| title_full | Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera |
| title_fullStr | Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera |
| title_full_unstemmed | Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera |
| title_short | Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera |
| title_sort | comparative evaluation of rbpt i elisa and cft for the diagnosis of brucellosis and pcr detection of brucella species from ethiopian sheep goats and cattle sera |
| topic | Brucella species Diagnosis CFT I-ELISA RBPT PCR |
| url | https://doi.org/10.1186/s12866-023-02962-2 |
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