Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results
False-positive serologic results (FPSRs) of brucellosis occur from time to time in various livestock with all the consequences (quarantine, compulsory slaughter, etc.) that follow true-positive laboratory results. A method based on the Polyacrylamide Gel Electrophoresis/Western Blot of a protein pan...
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MDPI AG
2025-03-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/13/3/574 |
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| author | Borbála Bányász József Antal Béla Dénes |
| author_facet | Borbála Bányász József Antal Béla Dénes |
| author_sort | Borbála Bányász |
| collection | DOAJ |
| description | False-positive serologic results (FPSRs) of brucellosis occur from time to time in various livestock with all the consequences (quarantine, compulsory slaughter, etc.) that follow true-positive laboratory results. A method based on the Polyacrylamide Gel Electrophoresis/Western Blot of a protein panel for resolving the FPSRs in the diagnosis of brucellosis was developed. Within the context of limited positive serum sample availability in Europe, the method successfully discriminates <i>Brucella</i>-positive sera from samples containing antibodies raised against infections caused by other Gram-negative bacteria causing FPSRs. An average CV% of 1.36 was determined for both repeatability and reproducibility for the whole separation mw range, and the test achieved 1.00 Diagnostic Sensitivity and 1.00 Diagnostic Specificity. The method with pre-prepared WB panels provides a rapid (less than 3 h), easily standardizable, and validatable alternative to existing confirmation methods. The whole WB process of the <i>Brucella</i> proteins and the subsequent densitometry can be accomplished with commercially available equipment, ready-to-use reagents, and open-source software, providing cost-effectiveness. The results of this study could attract broader attention, since molecular species in the 35.0–75.0 kDa range can serve as antigens in <i>Brucella</i> serology and the same fraction can be considered in the development of synthetic <i>Brucella</i> vaccines. |
| format | Article |
| id | doaj-art-0dae14fc59b3431eb35cdc7f2e5c2ccb |
| institution | Kabale University |
| issn | 2076-2607 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj-art-0dae14fc59b3431eb35cdc7f2e5c2ccb2025-08-20T03:43:10ZengMDPI AGMicroorganisms2076-26072025-03-0113357410.3390/microorganisms13030574Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test ResultsBorbála Bányász0József Antal1Béla Dénes2Institute of Isotopes Co., Ltd., 1121 Budapest, HungaryOmixon Biocomputing Ltd., 1117 Budapest, HungaryDepartment of Microbiology and Infectious Diseases, University of Veterinary Medicine Budapest, 1143 Budapest, HungaryFalse-positive serologic results (FPSRs) of brucellosis occur from time to time in various livestock with all the consequences (quarantine, compulsory slaughter, etc.) that follow true-positive laboratory results. A method based on the Polyacrylamide Gel Electrophoresis/Western Blot of a protein panel for resolving the FPSRs in the diagnosis of brucellosis was developed. Within the context of limited positive serum sample availability in Europe, the method successfully discriminates <i>Brucella</i>-positive sera from samples containing antibodies raised against infections caused by other Gram-negative bacteria causing FPSRs. An average CV% of 1.36 was determined for both repeatability and reproducibility for the whole separation mw range, and the test achieved 1.00 Diagnostic Sensitivity and 1.00 Diagnostic Specificity. The method with pre-prepared WB panels provides a rapid (less than 3 h), easily standardizable, and validatable alternative to existing confirmation methods. The whole WB process of the <i>Brucella</i> proteins and the subsequent densitometry can be accomplished with commercially available equipment, ready-to-use reagents, and open-source software, providing cost-effectiveness. The results of this study could attract broader attention, since molecular species in the 35.0–75.0 kDa range can serve as antigens in <i>Brucella</i> serology and the same fraction can be considered in the development of synthetic <i>Brucella</i> vaccines.https://www.mdpi.com/2076-2607/13/3/574brucellosisserologyfalse-positive serologic resultsprotein antigen panelwestern blotting |
| spellingShingle | Borbála Bányász József Antal Béla Dénes Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results Microorganisms brucellosis serology false-positive serologic results protein antigen panel western blotting |
| title | Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results |
| title_full | Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results |
| title_fullStr | Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results |
| title_full_unstemmed | Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results |
| title_short | Ghostbuster—A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results |
| title_sort | ghostbuster a western blot based panel method to resolve false positive brucellosis serology test results |
| topic | brucellosis serology false-positive serologic results protein antigen panel western blotting |
| url | https://www.mdpi.com/2076-2607/13/3/574 |
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