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Identification of Burkholderia mallei Isolates with Polymerase Chain Reaction-Restriction Fragment Length
F Abnaroodheleh1, N Mosavari2, S A Pourbakhsh2
1Veterinary Department, Science and Research Branch, Islamic Azad University, Tehran, Iran.
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.
Insights
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) effectively differentiates Burkholderia mallei, the cause of glanders, from Burkholderia pseudomallei. This molecular technique offers a precise and rapid diagnostic tool for identifying these zoonotic pathogens.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Zoonotic Diseases
Background:
- Glanders, caused by Burkholderia mallei, is a contagious zoonotic disease primarily affecting equids.
- Current epidemiological studies are limited by the unknown number of circulating strains and lack of effective prevention.
- Molecular techniques are crucial for accurate identification and tracking of bacterial strains in various settings.
Purpose of the Study:
- To evaluate the efficacy of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) for differentiating Burkholderia mallei and Burkholderia pseudomallei.
- To assess PCR-RFLP as a diagnostic tool for identifying field isolates of B. mallei.
Main Methods:
- Six field isolates and two laboratory strains of B. mallei and B. pseudomallei were cultured.
- Biochemical tests were performed for initial identification.
- DNA was extracted, and PCR-RFLP was conducted on the bacterial genomes. The Straus reaction was induced in guinea pigs.
Main Results:
- Biochemical assays confirmed the isolates as Burkholderia mallei.
- PCR-RFLP yielded a 650 bp product for B. mallei and 250/400 bp products for B. pseudomallei.
- The Straus reaction was observed in guinea pigs, indicated by scrotal swelling.
Conclusions:
- PCR-RFLP is a reliable differential diagnostic technique for B. mallei.
- This method accurately distinguishes between B. mallei and B. pseudomallei.
- PCR-RFLP provides rapid, precise, and sensitive detection of B. mallei.
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