Evaluation of indirect immunofluorescence assay for the diagnosis of bacterial agent of glanders

Jamooli, Shima; Mosavari, Nader; Assmar, Mahdi

year 10, Issue 3 (July - August 2016) Iran J Med Microbiol 2016, 10(3): 1-10 | Back to browse issues page

PMCID: 26289162

Mendeley

Zotero

RefWorks

Jamooli S, Mosavari N, Assmar M. Evaluation of indirect immunofluorescence assay for the diagnosis of bacterial agent of glanders. Iran J Med Microbiol 2016; 10 (3) :1-10
URL: http://ijmm.ir/article-1-478-en.html

Evaluation of indirect immunofluorescence assay for the diagnosis of bacterial agent of glanders

Shima Jamooli¹

, Nader Mosavari

², Mahdi Assmar³

1- Departeman of Microbiology, Faculty of Advance Sciences & Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran
2- Department of Tuberculin and Mallein Production and Research , Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran , n.mosavari@rvsri.ac.ir
3- Department of Parasitology, Pasteur Institute of Iran, Tehran, Iran

Abstract: (9850 Views)

Background and Aim: Glanders is one of the most dangerous and oldest communicable zoonotic diseases and causes many deaths in equines, carnivores, and humans. Because of the sudden increase of the rate of glanders in the Middle East and the use of its pathogen as a biological weapon, the detection of this disease is necessary by use of rapid diagnostic tests. This study was conducted to diagnose causative agent of glanders by immunofluorescence method in horses.

Materials and Methods: Burkholderia mallei strain was cultured in glycerinated gelose and bouin medium to produce mallein in the Department of Research and production of tuberculin and mallein in Razi Vaccine and Serum Research Institute, then identified using molecular techniques (Such as DNA extraction and PCR). A suspension was prepared and then heated to inactivate the bacteria. A bacterial smear was prepared and fixed on a slide. By injecting the microbial suspension to the horse, an antibody produced which was added to an anti-horse conjugated with fluorescein. The slide was examined using an immunofluorescent microscope.

Results: There were clearly luminous bacilli on the slide with positive serum, but dark bacilli were hardly observable on the slide with negative serum.

Conclusions: The results of observation of prepared slides from the positive and negative serum by ELISA, Malleination and CFT tests are match with the results of the immunofluorescence method and the validity of this method was confirmed. As a result, this method can be used for diagnosis causative agent of glanders.

Keywords: Glanders, Burkholderia mallei, Immunofluorecence

Full-Text [PDF 715 kb] (4742 Downloads)

Type of Study: Original Research Article | Subject: Medical Bacteriology
Received: 2015/09/10 | Accepted: 2016/04/15 | ePublished: 2016/10/16

References

1. Whitlock GC, Estes DM, Torres AG. Glanders: off to the races with Burkholderia mallei. FEMS Microbiol. Lett. 2007; 277 (2): 115–22. [PubMed]

2. Khaki P, Mosavari N, KhajehNasiri S, Emam M, Ahouran M, Hashemi S, et al.Glanders outbreak at Tehran Zoo, Iran. Iran J Med Microbiol. 2012; 4 (1): 3-7. [PubMed]

3. Vallat B, Caporale V. Manual of diagnostic tests and vaccines for terrestrial animals (mammals, birds and bees). 17nd ed. World Organisation for Animal Health (OIE); 2012; 2; Chapter 2.5.11. 913-20.

4. Schell MA, Ulrich RL, Ribot WJ, Brueggemann EE, Hines HB, Chen D, et al. Type VI secretion is amajor virulence determinant in Burkholderia mallei. Mol Microbiol. 2007; 64(6): 1466-85. [PubMed]

5. Tadjbakhsh H. Traditional methods used for controlling animal diseases in Iran. Rev Sci Tech. 1994; 13(2): 599-614. [PubMed]

6. Whitlock GC, Mark Estes D, Torres AG. Glanders: off to the races with Burkholderia mallei. FEMS Microbiol Lett. 2007; 277(2): 115-22. [PubMed]

7. Tajik A, Kazaemi A. The role of medical laboratory in diagnosis of NBC war lesions injuries.NEZAJA publication. 1nd ed; 2007: 89-94.

8. Lever MS, Nelson M, Ireland PI, Stagg AJ, Beedham RJ, Hall GA, et al. Experimentalaerogenic Burkholderia mallei (glanders) infection in the BALB/c mouse. J Med Microbiol. 2003; 52(12): 1109-15. [PubMed]

9. Blaha T. Applied Veterinary Epidemiology. 1nd ed. Tehran: Aeejh press; 1997.

10. Naureen A, Saqib M, Muhammad G, Hussain MH, Asi MN. Comparative evaluation of Rose Bengal plate agglutination test, mallein test, and some conventional serological tests for diagnosis of equine glanders. J Vet Diagn Invest. 2007; 19(4): 362-7. [PubMed]

11. Herr S, huchzermayer HF, Tebrugge LA, Williamson CC, Roos JA, Schiele GJ. The use of a single complement fixation technique in bovine brucellosis, Johne’s disease, dourine, equine piroplasmosis and Q fever serology. Onderstepoort J Vet Res. 1985; 52: 279-82. [PubMed]

12. Elschner MC, Scholz HC, Melzer F, Saqib M, Marten P, Rassbach A, et al. Use of a Western blot technique for the serodiagnosis of glanders. BMC vet res. 2011; 7(4): 1-6. [PubMed]

13. Van Zandt KE, Greer MT, Gelhaus HC. Glanders: an overview of infection in humans. Orphanet J Rare Dis. 2013; 8: 131-137. [PubMed]

14. Alibasoglu M, Yesildere T, Calislar T, Inal T, Calsikan U. Glanders outbreak in lions in the Istanbul zoological garden. Dtsch Tierarztl Wochenschr. 1986; 99: 57-63. [PubMed]

15. Naureen A, Saqib M, Ghulan M, Hussain MH, Asi MN. Comparative evaluation of Rose Bengal plate agglutination test, mallein test, and some conventional serological tests for diagnosis of equine glanders. J Vet Diagn Invest. 2007; 19: 362-367. [PubMed]

16. Al-Ani FK, Roberson J. Glanders in horses: A review of the literature. Vet Arh. 2007; 77(3): 203-18 [Article]

17. Akbarein H, Bahonar AR, Dabbagh Moghaddam A, Bolouki Z, Shokouh H. Glanders, A New Vision on an Old Biological Weapon. HBI J. 2012, 10(2): 143-62 [Article]

18. Tkachenko GA, Antonov VA, Zamaraev VS, Iliukhin VI. Identification of the causative agents of glanders and melioidosis by polymerase chain reaction. Mol Gen Mikrobiol Virusol. 2003;(3): 18-22. [PubMed]

19. Filho MBC, Ramos RM, Fonseca AA, Orzil LL, Sales ML, Santana VLA , et al. Development and validation of a method for purification of mallein for the diagnosis of glanders in equines. BMC Vet Res. 2012; 8:154-63. [Article]

20. Poppert S, Elschner M, Tannich E, Vergenes LM, Hagen R. Identification of Burkholderia Pseudomallei and B.malleiby Fluorescence in situe Hybridization. Int J Med Microbiol. 2011; 301(7): 585-90.