Original Article

Genomic pattern analysis of Burkholderia mallei field isolates by pulsed-field gel electrophoresis (PFGE) discriminatory typing


Background and Objectives: Glanders is a serious zoonotic disease caused by Burkholderia mallei. Prevention, control, and treatment strategies of glanders are prerequisites for microbial source tracking. The present study was aimed to analyze the genomic pattern of B. mallei Iranian field isolates by pulsed-field gel electrophoresis (PFGE) typing.
Materials and Methods: B. mallei isolates were aerobically cultured in nutrient broth/agar supplemented with glycerol 4% for 48 h at 37°C. API 20NE identification system was used for the biochemical characterization. Genomic DNA of bacterial isolates was extracted using OIE-recommended protocol. Molecular identification of bacterial isolates was done based on amplification of BimA and IS407-flip genes. PFGE was applied to prepare the genomic pattern of B. mallei isolates. The guinea pig was used as a suitable model for studying the histopathological characterization of B. mallei.
Results: In both enzymatic digestion patterns by using Af1II and VspI, we found three different clonal types; І) PFGE type of B. mallei Razi 325 strain, ІІ) PFGE type of Tiger, Kordan, and Oshnavieh strains, and ІІІ) PFGE type of Semirom strain. B. mallei Razi 325 was categorized as unrelated strain which was belonged to the different cluster differing more than four bands.
Conclusion: PFGE showed more discriminatory power and considerable reproducibility for molecular typing of B. mallei strains in our study. It is standardized the approaches for outbreak detection, pathogen phylogeny, molecular epidemiology, and population studies.

1. Nguyen HN, Smith ME, Hayoun MA (2021). Glanders and melioidosis. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing.
2. Khan I, Wieler LH, Melzer F, Elschner MC, Muhammad G, Ali S, et al. Glanders in animals: a review on epidemiology, clinical presentation, diagnosis and countermeasures. Transbound Emerg Dis 2013;60:204-221.
3. Van Zandt KE, Greer MT, Gelhaus HC. Glanders: an overview of infection in humans. Orphanet J Rare Dis 2013;8:131.
4. Inglis TJJ, Merritt AJ (2015). Burkholderia pseudomallei and Burkholderia mallei. In: Molecular Medical Microbiology. Ed(s), YW Tang, M Sussman, D Liu, I Poxton, J Schwartzman. Academic Press, 2nd ed. Crawley, WA, Australia, pp. 769-791.
5. Schutzer SE, Schlater LR, Ronning CM, DeShazer D, Luft BJ, Dunn JJ, et al. Characterization of clinically-attenuated Burkholderia mallei by whole genome sequencing: candidate strain for exclusion from select agent lists. PLoS One 2008;3(4):e2058.
6. Yazdansetad S, Mosavari N, Tadayon K, Mehregan I. Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm. Iran J Microbiol 2019;11:232-238.
7. Khaki P, Mosavari N, Khajeh NS, Emam M, Ahouran M, Hashemi S, et al. Glanders outbreak at Tehran zoo, Iran. Iran J Microbiol 2012;4:3-7.
8. OIE World Organization for Animal Health [OIE]. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals 2019.
9. Ulrich RL, Ulrich MP, Schell MA, Kim HS, DeShazer D. Development of a polymerase chain reaction assay for the specific identification of Burkholderia mallei and differentiation from Burkholderia pseudomallei and other closely related Burkholderiaceae. Diagn Microbiol Infect Dis 2006;55:37-45.
10. Scholz HC, Joseph M, Tomaso H, Al Dahouk S, Witte A, Kinne J, et al. Detection of the reemerging agent Burkholderia mallei in a recent outbreak of glanders in the United Arab Emirates by a newly developed fliP-based polymerase chain reaction assay. Diagn Microbiol Infect Dis 2006;54:241-247.
11. Bauernfeind A, Roller C, Meyer D, Jungwirth R, Schneider I. Molecular procedure for rapid detection of Burkholderia mallei and Burkholderia pseudomallei. J Clin Microbiol 1998;36:2737-2741.
12. Lowe W, March JK, Bunnell AJ, O’Neill KL, Robison RA. PCR-based methodologies used to detect and differentiate the Burkholderia pseudomallei complex: B. pseudomallei, B. mallei, and B. thailandensis. Curr Issues Mol Biol 2014;16:23-54.
13. Lee MA, Wang D, Yap EH. Detection and differentiation of Burkholderia pseudomallei, Burkholderia mallei and Burkholderia thailandensis by multiplex PCR. FEMS Immunol Med Microbiol 2005;43:413-417.
14. Mirzai S, Safi S, Mossavari N, Afshar D, Bolourchian M. Development of a loop-mediated isothermal amplification assay for rapid detection of Burkholderia mallei. Cell Mol Biol (Noisy-le-grand) 2016;62:32-36.
15. Lowe CW, Satterfield BA, Nelson DB, Thiriot JD, Heder MJ, March JK, et al. A quadruplex real-time PCR assay for the rapid detection and differentiation of the most relevant members of the B. pseudomallei complex: B. mallei, B. pseudomallei, and B. thailandensis. PLoS One 2016;11(10):e0164006.
16. Stevens JM, Ulrich RL, Taylor LA, Wood MW, Deshazer D, Stevens MP, et al. Actin-binding proteins from Burkholderia mallei and Burkholderia thailandensis can functionally compensate for the actin-based motility defect of a Burkholderia pseudomallei bimA mutant. J Bacteriol 2005;187:7857-7862.
17. Stevens MP, Stevens JM, Jeng RL, Taylor LA, Wood MW, Hawes P, et al. Identification of a bacterial factor required for actin-based motility of Burkholderia pseudomallei. Mol Microbiol 2005;56:40-53.
18. Jakupciak JP, Wells JM, Karalus RJ, Pawlowski DR, Lin JS, Feldman AB. Population-sequencing as a biomarker of Burkholderia mallei and Burkholderia pseudomallei evolution through microbial forensic analysis. J Nucleic Acids 2013;2013:801505.
19. Godoy D, Randle G, Simpson AJ, Aanensen DM, Pitt TL, Kinoshita R, et al. Multilocus sequence typing and evolutionary relationships among the causative agents of melioidosis and glanders, Burkholderia pseudomallei and Burkholderia mallei. J Clin Microbiol 2003;41:2068-2079.
20. Gilmour MW, Graham M, Reimer A, Van Domselaar G. Public health genomics and the new molecular epidemiology of bacterial pathogens. Public Health Genomics 2013;16:25-30.
21. Ko WC, Cheung BM, Tang HJ, Shih HI, Lau YJ, Wang LR, et al. Melioidosis outbreak after typhoon, southern Taiwan. Emerg Infect Dis 2007;13:896-898.
22. Chua KH, See KH, Thong KL, Puthucheary SD. SpeI restriction enzyme displays greater discriminatory power than XbaI enzyme does in a pulsed-field gel electrophoresis study on 146 clinical Burkholderia pseudomallei isolates. Jpn J Infect Dis 2011;64:228-233.
23. Majerczyk CD, Brittnacher MJ, Jacobs MA, Armour CD, Radey MC, Bunt R, et al. Cross-species comparison of the Burkholderia pseudomallei, Burkholderia thailandensis, and Burkholderia mallei quorum-sensing regulons. J Bacteriol 2014;196:3862-3871.
24. Chantratita N, Vesaratchavest M, Wuthiekanun V, Tiyawisutsri R, Ulziitogtokh T, Akcay E, et al. Pulsed-field gel electrophoresis as a discriminatory typing technique for the biothreat agent Burkholderia mallei. Am J Trop Med Hyg 2006;74:345-347.
IssueVol 13 No 5 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v13i5.7419
Burkholderia mallei; Pulsed-field gel electrophoresis; Glanders; Zoonoses; Biological warfare

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Dashtipour S, Tadayon K, Yazdansetad S, Mosavari N, Keshavarz R. Genomic pattern analysis of Burkholderia mallei field isolates by pulsed-field gel electrophoresis (PFGE) discriminatory typing. Iran J Microbiol. 2021;13(5):574-582.