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Designing of Western Blot Technique for Glanders Diagnosing in Iran
Nafiseh Shakibamehr1, Nader Mosavari2, Nasser Harzandi1
1Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran.
Abstract:
Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic disease in its early stages. Some methods such as the complement fixation test (CFT) cause some problems for veterinary authorities and financial losses to animal owners due to false-positive results. The mallein test requires appropriate laboratory equipment and skilled personnel. To quickly and accurately diagnose the disease, especially in areas where animals cannot be kept, new methods (such as the Western blot test [WBT]) should be used to identify the disease. This study designed and optimized the Western blot (immunoblot) test using sera from 84 glanderous equids, and the sensitivity and specificity of ELISA and CFT were compared with the WBT. ELISA tests are based on B. mallei antigens whereas a purified lipopolysaccharide-containing B. mallei antigen is used in the WBT. The sensitivity and specificity of the tests were estimated using the cut-off values recommended by the test developers. The WBT and ELISA were significantly more specific than the CFT. The ELISA based on B. mallei antigens was significantly less sensitive than the CFT. Given their comparable sensitivities and specificities, the CFT (95.7%, 98.5%), the WBT (95%, 100%) and the ELISA (85%, 100%) should be further developed. The CFT is still the prescribed technique for serological investigation of equids for trade purposes to certify individual animals without glanders. Therefore, more efforts should be made to further improve and optimize the WBT and ELISA tests.
Insights
Diagnosing glanders in equids is challenging. The Western blot test (WBT) and ELISA show promise for accurate diagnosis, offering higher specificity than the complement fixation test (CFT).
Area of Science:
- Veterinary diagnostics
- Zoonotic disease research
- Microbiology
Background:
- Glanders, caused by *Burkholderia mallei*, is a difficult-to-diagnose zoonotic disease.
- Current diagnostic methods like complement fixation test (CFT) and mallein test have limitations, including false positives and resource requirements.
- Accurate and rapid diagnostic tools are crucial, especially for animals that cannot be housed in specific facilities.
Purpose of the Study:
- To design and optimize the Western blot (immunoblot) test (WBT) for diagnosing glanders in equids.
- To compare the sensitivity and specificity of WBT with ELISA and CFT.
- To evaluate the diagnostic performance of WBT, ELISA, and CFT for glanders detection.
Main Methods:
- The study involved sera from 84 equids with glanders.
- A Western blot (immunoblot) test was designed and optimized using *B. mallei* antigens.
- Sensitivity and specificity of WBT, ELISA (using *B. mallei* antigens), and CFT were compared using established cut-off values.
Main Results:
- Western blot test (WBT) and ELISA demonstrated significantly higher specificity compared to CFT.
- ELISA showed significantly lower sensitivity than CFT.
- The diagnostic performance was CFT (95.7% sensitivity, 98.5% specificity), WBT (95% sensitivity, 100% specificity), and ELISA (85% sensitivity, 100% specificity).
Conclusions:
- WBT and ELISA are highly specific diagnostic methods for glanders, outperforming CFT in specificity.
- Further development and optimization of WBT and ELISA are recommended for improved sensitivity and broader application.
- Despite limitations, CFT remains the standard for serological testing in equids for trade purposes.
