Volume 13, Issue 1 (Jan-Feb 2019)                   mljgoums 2019, 13(1): 33-36 | Back to browse issues page


XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Rezaei E, Khaki P, Moradi Bidhendi S, Noofeli M. Identification of Pathogenic Leptospiral Serovars by Detection of the ompl37 Gene Using PCR. mljgoums 2019; 13 (1) :33-36
URL: http://mlj.goums.ac.ir/article-1-1161-en.html
1- Department of Microbiology, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
2- Department of Microbiology, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran , P.khaki@rvsri.ac.ir
3- Department of human bacterial vaccines, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
Abstract:   (7036 Views)
ABSTRACT
            Background and Objectives: Leptospirosis is a widespread zoonotic disease that is transmitted directly or indirectly from animals to humans. Humans mainly acquire pathogenic leptospires through mucosal or percutaneous exposure to environment contaminated with urine from an infected animal. We aimed to identify pathogenic leptospiral serovars by detection of the ompL37 gene using polymerase chain reaction (PCR).
            Methods: Sixteen pathogenic leptospiral serovars and a saprophytic serovar, L. biflexa were cultured in modified semisolid Ellinghausen-McCullough-Johnson-Harris medium containing 5% rabbit serum. Genomic DNA extraction was done using the phenol-chlorophorm method. The ompL37 gene was amplified using specific primers. PCR products were analyzed by agarose gel electrophoresis.
            Results: The ompL37 gene was amplified only in the pathogenic leptospiral serovars. We detected no amplified fragment for the saprophytic serovar.
Conclusion: Leptospirosis may be confused with other infectious diseases, and therefore, its early and accurate diagnosis is crucial. We showed that molecular detection of pathogenic leptospires based on the ompL37 gene could be used for laboratory diagnosis of leptospirosis.
            Keywords: Leptospirosis, PCR, ompl37 Gene, Pathogenic Leptospires.
Full-Text [PDF 505 kb]   (1485 Downloads)    
Research Article: Original Paper | Subject: Laboratory Sciences
Received: 2018/12/22 | Accepted: 2018/12/22 | Published: 2018/12/22 | ePublished: 2018/12/22

References
1. Haake DA, Levett PN. Leptospirosis in humans. Leptospira and leptospirosis: Springer; 2015. p. 65-97.
2. Mullan S, Panwala TH. Polymerase chain reaction: an important tool for early diagnosis of leptospirosis cases. Journal of clinical and diagnostic research: JCDR. 2016; 10(12): DC08. [DOI:10.7860/JCDR/2016/22462.9010]
3. Monahan A, Miller I, Nally J. Leptospirosis: risks during recreational activities. Journal of applied microbiology. 2009; 107(3): 707-16. [DOI:10.1111/j.1365-2672.2009.04220.x]
4. Plank R, Dean D. Overview of the epidemiology, microbiology, and pathogenesis of Leptospira spp. in humans. Microbes and infection. 2000; 2(10): 1265-76. [DOI:10.1016/S1286-4579(00)01280-6]
5. Picardeau M, Bertherat E, Jancloes M, Skouloudis AN, Durski K, Hartskeerl RA. Rapid tests for diagnosis of leptospirosis: current tools and emerging technologies. Diagnostic microbiology and infectious disease. 2014; 78(1): 1-8. Diagn Microbiol Infect Dis. 2014; 78(1): 1-8. doi: 10.1016/j.diagmicrobio.2013.09.012. [DOI:10.1016/j.diagmicrobio.2013.09.012]
6. Lilenbaum W, Varges R, Brandão F, Cortez A, De Souza S, Brandão P, et al. Detection of Leptospira spp. in semen and vaginal fluids of goats and sheep by polymerase chain reaction. Theriogenology. 2008; 69(7): 837-42. doi: 10.1016/j.theriogenology.2007.10.027. [DOI:10.1016/j.theriogenology.2007.10.027]
7. Hoseinpur R, Khaki P, Noofeli M, Bidhendi S. Molecular detection of pathogenic leptospiral serovars by PCR, based on lipL21 gene. Archives of Razi Institute. 2016; 70(4): 223-7.
8. Azizi S, Kheirandish R, Rahimi E. Comparison of polymerase chain reaction and Warthin-Starry techniques to detect Leptospira spp. in kidneys of slaughtered cattle. Onderstepoort J Vet Res. 2014 ; 81(1): e1-e6. doi: 10.4102/ojvr.v81i1.821. [DOI:10.4102/ojvr.v81i1.821]
9. Oliveira TL, Grassmann AA, Schuch RA, Neto ACPS, Mendonça M, Hartwig DD, et al. Evaluation of the Leptospira interrogans outer membrane protein OmpL37 as a vaccine candidate. PloS one. 2015;10(11):e0142821. [DOI:10.1371/journal.pone.0142821]
10. Pinne M, Choy HA, Haake DA. The OmpL37 surface-exposed protein is expressed by pathogenic Leptospira during infection and binds skin and vascular elastin. PLoS Negl Trop Dis. 2010; 4(9): e815. doi: 10.1371/journal.pntd.0000815. [DOI:10.1371/journal.pntd.0000815]
11. Sambrook, J., & Russell, D. W. Rapid isolation of yeast DNA. Cold Spring Harbor Protocols. 2002; (1), pdb. doi:10.1101/pdb.prot4039. [DOI:10.1101/pdb.prot4039]
12. Langston CE, Heuter KJ. Leptospirosis: A re-emerging zoonotic disease. Vet Clin North Am Small Anim Pract. 2003; 33(4): 791-807. [DOI:10.1016/S0195-5616(03)00026-3]
13. Schneider MC, Jancloes M, Buss DF, Aldighieri S, Bertherat E, Najera P, et al. Leptospirosis: a silent epidemic disease. Int J Environ Res Public Health. 2013; 10(12): 7229-7234. doi: 10.3390/ijerph10127229. [DOI:10.3390/ijerph10127229]
14. Fraga T, Barbosa A, Isaac L. Leptospirosis: aspects of innate immunity, immunopathogenesis and immune evasion from the complement system. Scand J Immunol. 2011; 73(5): 408-19. doi: 10.1111/j.1365-3083.2010.02505.x. [DOI:10.1111/j.1365-3083.2010.02505.x]
15. Helmerhorst H, Van Tol E, Tuinman P, De Vries P, Hartskeerl R, Grobusch M, et al. Severe pulmonary manifestation of leptospirosis. Neth J Med. 2012; 70(5): 215-21.
16. Majed Z, Bellenger E, Postic D, Pourcel C, Baranton G, Picardeau M. Identification of variable-number tandem-repeat loci in Leptospira interrogans sensu stricto. J Clin Microbiol. 2005; 43(2): 539-45. [DOI:10.1128/JCM.43.2.539-545.2005]
17. Slack A, Symonds M, Dohnt M, Smythe L. An improved multiple-locus variable number of tandem repeats analysis for Leptospira interrogans serovar Australis: a comparison with fluorescent amplified fragment length polymorphism analysis and its use to redefine the molecular epidemiology of this serovar in Queensland, Australia. J Med Microbiol. 2006; 55(Pt 11): 1549-57. [DOI:10.1099/jmm.0.46779-0]
18. Slack AT, Dohnt MF, Symonds ML, Smythe LD. Development of a multiple-locus variable number of tandem repeat analysis (MLVA) for Leptospira interrogans and its application to Leptospira interrogans serovar Australis isolates from Far North Queensland, Australia. Annals of clinical microbiology and antimicrobials. 2005;4(1):10. [DOI:10.1186/1476-0711-4-10]

Add your comments about this article : Your username or Email:
CAPTCHA

Send email to the article author


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2007 All Rights Reserved | Medical Laboratory Journal

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.