year 16, Issue 5 (September - October 2022)
Iran J Med Microbiol 2022, 16(5): 392-398 |
Back to browse issues page
1- Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
2- Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran , Khakipejvak53@gmail.com
3- Department of Research and Development, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
2- Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran , Khakipejvak53@gmail.com
3- Department of Research and Development, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
Abstract: (1810 Views)
Background and Aim: Leptospirosis is one of the most common zoonotic diseases worldwide, occurring mostly in tropical, subtropical, temperate, and humid regions with heavy rainfall. It is important to diagnose this condition correctly and promptly. Loa22 is an outer membrane protein exposed to the surface in some Leptospira serovars. The purpose of this study was to determine the presence of the gene encoding Loa22 protein in Leptospira interrogans serovars.
Materials and Methods: The present study was conducted on 23 pathogenic leptospira serovars and two non-pathogenic leptospira serovars. These serovars were prepared from the Reference Laboratory for Leptospira, Department of Microbiology, Razi Vaccine, and Serum Research Institute, Karaj, Iran. After genomic DNA extraction using the standard phenol-chloroform method, loa22 gene was amplified by specific primers.
Results: PCR was performed on the loa22 gene by producing a 671-bp fragment. The results showed that the loa22 gene was present in all 23 pathogenic leptospira serovars but not in the non-pathogenic L. biflexa. The specificity of tested primers was confirmed as well.
Conclusion: The loa22 gene is a specific gene for pathogenic leptospira serovars that is not found in saprophytic serovars, so it is suggested that this gene be used to detect leptospira pathogenic serovars.
Type of Study: Original Research Article |
Subject:
Zoonoses Research
Received: 2021/10/18 | Accepted: 2022/05/28 | ePublished: 2022/08/8
Received: 2021/10/18 | Accepted: 2022/05/28 | ePublished: 2022/08/8
References
1. Soo ZMP, Khan NA, Siddiqui R. Leptospirosis: Increasing importance in developing countries. Acta trop. 2020;201:105183. [DOI:10.1016/j.actatropica.2019.105183] [PMID]
2. Picardeau M. Diagnosis and epidemiology of Leptospirosis. Med Mal Infect. 2013;43(1):1-9. [DOI:10.1016/j.medmal.2012.11.005] [PMID]
3. Musso D, La Scola B. Laboratory diagnosis of Leptospirosis: a challenge. J Microbiol Immunol Infect. 2013;46(4):245-52. [DOI:10.1016/j.jmii.2013.03.001] [PMID]
4. Victoriano AFB, Smythe LD, Gloriani-Barzaga N, Cavinta LL, Kasai T, Limpakarnjanarat K, et al. Leptospirosis in the Asia Pacific region. BMC Infect Dis. 2009;9(1):1-9. [DOI:10.1186/1471-2334-9-147] [PMID] [PMCID]
5. Narkkul U, Thaipadungpanit J, Srisawat N, Rudge JW, Thongdee M, Pawarana R, et al. Human, animal, water source interactions and Leptospirosis in Thailand. Sci Rep. 2021;11(1):1-13. [DOI:10.1038/s41598-021-82290-5] [PMID] [PMCID]
6. Babamahmodi F, Motamed N, Mahdavi M, Nickhah F, Qavi Bonyeh K. Seroepidemiological study of Leptospirosis in Ghaemshahr Mazandaran province-Iran, Sept-Oct 2004. J Maz Univ Med Sci. 2006;16(53):51-6.
7. Bajani MD, Ashford DA, Bragg SL, Woods CW, Aye T, Spiegel RA, et al. Evaluation of four commercially available rapid serologic tests for diagnosis of Leptospirosis. J Clin Microbiol. 2003;41(2):803-9. [DOI:10.1128/JCM.41.2.803-809.2003] [PMID] [PMCID]
8. Hamond C, Martins G, Loureiro A, Pestana C, Lawson-Ferreira R, Medeiros M, et al. Urinary PCR as an increasingly useful tool for an accurate diagnosis of Leptospirosis in livestock. Vet Res Commun. 2014;38(1):81-5. [DOI:10.1007/s11259-013-9582-x] [PMID]
9. Perez J, Goarant C. Rapid Leptospira identification by direct sequencing of the diagnostic PCR products in New Caledonia. BMC Microbiol. 2010;10(1):1-11. [DOI:10.1186/1471-2180-10-325] [PMID] [PMCID]
10. Di Azevedo M, Lilenbaum W. An overview on the molecular diagnosis of animal leptospirosis. Lett Appl Microbiol. 2021;72(5):496-508. [DOI:10.1111/lam.13442] [PMID]
11. Riediger IN, Stoddard RA, Ribeiro GS, Nakatani SM, Moreira SD, Skraba I, et al. Rapid, actionable diagnosis of urban epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay. PLoS neglected tropical diseases. 2017;11(9):e0005940. [DOI:10.1371/journal.pntd.0005940] [PMID] [PMCID]
12. Dezhbord MA KP, Esmaili Zadeh M, Salehi B, Moradi Bidhendi S, Khodaverdi K, et al. Differentiation of pathogenic serovars from non-pathogenic Leptospira by PCR based on ompLI gene and design of a positive control to optimize this diagnostic test. IJMM. 2012;6(3):45-51.
13. Murray GL, Srikram A, Henry R, Puapairoj A, Sermswan RW, Adler B. Leptospira interrogans requires heme oxygenase for disease pathogenesis. Microbes Infect. 2009;11(2):311-4. [DOI:10.1016/j.micinf.2008.11.014] [PMID]
14. Cullen PA, Haake DA, Adler B. Outer membrane proteins of pathogenic spirochetes. FEMS Microbiol Rev. 2004;28(3):291-318. [DOI:10.1016/j.femsre.2003.10.004] [PMID] [PMCID]
15. Li X, Bao L, Hu C, Xie Y, Yan J, Zhang H. Molecular cloning and expression in E. coli of the surface-exposed lipoprotein LipL41 gene of Leptospira lai. Hua xi yi ke da xue xue bao= Journal of West China University of Medical Sciences= Huaxi Yike Daxue Xuebao. 2001;32(3):341-3, 448.
16. Podgoršek D, Ružić-Sabljić E, Logar M, Pavlović A, Remec T, Baklan Z, et al. Evaluation of real-time PCR targeting the lipL32 gene for diagnosis of Leptospira infection. BMC Microbiol. 2020;20 (1):1-9. [DOI:10.1186/s12866-020-01744-4] [PMID] [PMCID]
17. Palaniappan RU, Chang Y-F, Chang C-F, Pan M, Yang C, Harpending P, et al. Evaluation of lig-based conventional and real time PCR for the detection of pathogenic leptospires. Mol Cell Probes. 2005;19(2):111-7. [DOI:10.1016/j.mcp.2004.10.002] [PMID]
18. Bal A, Gravekamp C, Hartskeerl R, De Meza-Brewster J, Korver H, Terpstra W. Detection of leptospires in urine by PCR for early diagnosis of Leptospirosis. J Clin Microbiol. 1994;32(8):1894-8. [DOI:10.1128/jcm.32.8.1894-1898.1994] [PMID] [PMCID]
19. Umthong S, Buaklin A, Jacquet A, Sangjun N, Kerdkaew R, Patarakul K, et al. Immunogenicity of a DNA and recombinant protein vaccine combining LipL32 and Loa22 for Leptospirosis using chitosan as a delivery system. J Microbiol Biotechnol. 2015;25(4):526-36. [DOI:10.4014/jmb.1408.08007] [PMID]
20. Nally JE, Whitelegge JP, Bassilian S, Blanco DR, Lovett MA. Characterization of the outer membrane proteome of Leptospira interrogans expressed during acute lethal infection. Infect Immun. 2007;75(2):766-73. [DOI:10.1128/IAI.00741-06] [PMID] [PMCID]
21. Hsu S-H, Chang M-Y, Ko Y-C, Chou L-F, Tian Y-C, Hung C-C, et al. Peptidoglycan Mediates Loa22 and Toll-like Receptor 2 Interactions in Pathogenic Leptospira. bioRxiv. 2019:520288. [DOI:10.1101/520288]
22. Sabri A, Khairani-Bejo S, Zunita Z, Hassan L. Molecular detection of Leptospira sp. in cattle and goats in Kelantan, Malaysia after a massive flood using multiplex polymerase chain reaction. Trop Biomed. 2019;36:165-71.
23. Darian EK, Forghanifard MM, Bidhendi SM, Chang Y-F, Yahaghi E, Esmaelizad M, et al. Cloning and sequence analysis of LipL32, a surface-exposed lipoprotein of pathogenic Leptospira spp. Iran Red Crescent Med J. 2013;15(11). [DOI:10.5812/ircmj.8793] [PMID] [PMCID]
24. Dezhbord M, Esmaelizad M, Khaki P, Fotohi F, Moghaddam AZ. Molecular identification of the ompL1 gene within Leptospira interrogans standard serovars. J Infect Dev Ctrie. 2014;8(06) :688-93. [DOI:10.3855/jidc.3174] [PMID]
25. Natarajaseenivasan K, Vijayachari P, Sharma S, Sugunan A, Sehgal S. Phenotypic & genotypic conservation of ompL1 & lipL41 among leptospiral isolates of Andaman Islands. Indian J Med Res. 2005;122(4):343.
26. Koizumi N, Watanabe H. Molecular cloning and characterization of a novel leptospiral lipoprotein with OmpA domain. FEMS Microbiol Lett. 2003;226(2):215-9. [DOI:10.1016/S0378-1097(03)00619-0]
27. Golsha R, Khodabakhshi B, RAHNAMA A. eptospirosis in Golestan province in Iran (Reports of twelve cases). J Gorgan Univ Med Sci. 2007;9(2):76-80.
28. Varadarajan MT, Gopalakrishnan R, Govindan B, Rajendiran A, Kathaperumal K. Virulence Gene Loa22-The Molecular Diagnostic Beacon of Canine Leptospirosis. Int j chem environ biol sci. 2015;3(1):21-4.
29. Meenambigai T, Anupriya R, Balakrishnan G, Ravikumar G, Ganesan P. Sensitivity of Virulence Gene Loa22 in a Multiplex PCR for Detection of Canine Leptospirosis. Int J Trend Res Dev. 2016;3(6):573-576.
30. Haake DA, Zückert WR. The leptospiral outer membrane. Leptospira and Leptospirosis. 2015;2015:187-221. [DOI:10.1007/978-3-662-45059-8_8] [PMID] [PMCID]
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |