Identification of Burkholderia mallei Isolates with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.
Abstract: Burkholderia mallei is the main cause of glanders as a dangerous contagious zoonosis disease that is mostly observed in single-hoofed animals, especially horses. Modern molecular techniques have been recently employed to improve epidemiology for identifying and searching for strains of this bacterium at different times and locations. Due to the unknown number of circulating strains and lack of preventive methods, glanders is still observed in the form of epidemics. The present study aimed to evaluate six field isolates plus two laboratory strains of Borkolderia mallei and Burkholderia pseudomallei using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. All the isolates and strains were microbially cultured in the glycerol nutrient and glycerol agar media. The individually grown colonies of the bacterium were used in the biochemical tests. The DNA of isolates was extracted by boiling, and the PCR-RFLP test was conducted on their genome. Finally, the bacterium was injected into guinea pigs to induce the Straus reaction. The biochemical assays (or bioassays) confirmed the isolates as Burkholderia mallei. The PCR-RFLP assay demonstrated a product for Burkholderia mallei with a length of 650 bp. Nevertheless, 250 and 400 bp were produced for Burkholderia pseudomallei. The swollen scrotum pointed to the occurrence of the Straus reaction. The PCR-RFLP is a proper differential diagnosis technique for B. mallei; moreover, it is a suitable method for differentiating between Burkholderia mallei and Burkholderia pseudomallei. This technique can detect Burkholderia mallei in a short time with high precision and sensitivity.
Publication Date: 2023-08-31 PubMed ID: 38226390PubMed Central: PMC10787924DOI: 10.32592/ARI.2023.78.4.1305Google Scholar: Lookup
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- Journal Article
Summary
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The research focuses on the application of molecular techniques, specifically PCR-RFLP, to identify strains of Burkholderia mallei, a bacterium that causes a contagious zoonosis disease known as glanders.
Objective and methodology
- The study aimed to assess the effectiveness of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in identifying strains of Burkholderia mallei and distinguishing them from other bacteria species.
- Six field isolates and two laboratory strains of Burkholderia mallei and Burkholderia pseudomallei were evaluated.
- The bacterial strains and isolates were cultured in glycerol nutrient and glycerol agar media to facilitate growth.
- Biochemical tests were performed on the individually grown bacterial colonies.
- The DNA of the isolates was extracted using a boiling method, and the PCR-RFLP test was performed on the extracted genome.
- Finally, the bacterium was injected into guinea pigs to provoke the Straus reaction, a telltale symptom of glanders.
Key findings
- The biochemical tests confirmed the identity of the isolates as Burkholderia mallei.
- In the PCR-RFLP test, a product length of 650 base pairs on the gel was indicative of Burkholderia mallei, whereas lengths of 250 and 400 base pairs represented Burkholderia pseudomallei.
- The occurrence of the Straus reaction in the guinea pigs (evidenced by a swollen scrotum) further substantiated the presence of Burkholderia mallei.
Conclusions and implications
- The researchers concluded that the PCR-RFLP method is an effective diagnostic tool for differentiating the strains of Burkholderia mallei from other bacteria, highlighting its sensitivity, precision, and rapid detection capabilities.
- This study’s findings provide valuable insights for clinical and public health usage, particularly in the rapid identification, prevention, and outbreak control of glanders disease.
Cite This Article
APA
Abnaroodheleh F, Mosavari N, Pourbakhsh SA, Tadayon K, Jamshidian M.
(2023).
Identification of Burkholderia mallei Isolates with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.
Arch Razi Inst, 78(4), 1305-1312.
https://doi.org/10.32592/ARI.2023.78.4.1305 Publication
Researcher Affiliations
- Veterinary Department, Science and Research Branch, Islamic Azad University, Tehran, Iran.
- Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
- Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
- Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
- Veterinary Department, Science and Research Branch, Islamic Azad University, Tehran, Iran.
MeSH Terms
- Horses / genetics
- Animals
- Guinea Pigs
- Burkholderia mallei / genetics
- Glanders / diagnosis
- Glanders / microbiology
- Polymorphism, Restriction Fragment Length
- Glycerol
- Burkholderia pseudomallei / genetics
- Polymerase Chain Reaction / veterinary
- Polymerase Chain Reaction / methods
- Horse Diseases
Conflict of Interest Statement
The authors declare that they have no conflict of interest.
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Citations
This article has been cited 1 times.- Sukmanadi M, Khairullah AR, Wardhani BWK, Mustofa I, Aliyah SH, Moses IB, Ahmad RZ, Khalisa AT, Pratama BP, Kusala MKJ, Kurniasih DAA, Akintunde AO, Fauziah I, Wibowo S, Furqoni AH, Fauzia KA, Melati I, Kurniawan M'. Glanders: Historical military use and potential bioterrorism concern. Open Vet J 2025 Sep;15(9):3912-3930.
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