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Application of polymerase chain reaction (PCR) methods in distinguishing pathogenic and saprophytic Leptospira from the traditional market environment in Denpasar City

Abstract

Background: Leptospirosis disease is still a health problem in Indonesia. The number of leptospirosis cases and deaths in Indonesia tends to increase. Indirect transmission of Leptospira bacteria can occur through poor environmental sanitation polluted by rat urine has the potential to cause leptospirosis. Traditional markets are good places for rats to breed. The purpose of this study was to detect the presence of contamination by pathogenic and saprophytic Leptospira bacteria in the environment around traditional markets in Denpasar City by polymerase chain reaction (PCR) method using three specific primers.

Methods: The sample is water from the market environment, taken at 19 location points from eight traditional markets in Denpasar City. After being homogenized, the sample water is filtered. Isolation of specific genes from samples by the PCR method was performed to differentiate pathogenic and saprophytic leptospira using three specific primers designed from the 16S rRNA gene. Starting from the DNA extraction stage, amplification by PCR, and detection of PCR product DNA by electrophoresis.

Results: This study found that from 19 water sampling locations, 5/19 (26.3%) point locations found specific DNA genes for pathogenic Leptospira bacteria. 10/19 (52.6%) point locations found specific genes for saprophytic Leptospira bacteria DNA, and 4/19 (21.1%) point locations found no specific genes for pathogenic or saprophytic Leptospira bacteria.

Conclusion: PCR examination using a combination of three primers could distinguish the presence of pathogenic Leptospira, saprophytic Leptospira, and the absence of pathogenic and saprophytic Leptospira simultaneously from traditional market environments in Denpasar City.

Section

References

  1. Ministry of Health of the Republic of Indonesia. Leptospirosis Control Technical Instructions, 3rd Printing. Jakarta: Directorate General of Disease Prevention and Control; 2017.
  2. Ariania N, Wahyono TYM. Faktor – faktor yang mempengaruhi kejadian Leptospirosis di 2 Kabupaten lokasi surveilans sentinel Leptospirosis Provinsi Banten tahun 2017 – 2019 Artikel Penelitian. Jurnal Epidemiologi Kesehatan Indonesia. 2020;4(2):57-63.
  3. Grillova´ L, Angermeier H, Levy M, Giard M, Lastère S, Picardeau M. Circulating genotypes of Leptospira in French Polynesia : An 9- year molecular epidemiology surveillance follow-up study. PLoS Negl Trop Dis. 2020;14(9):e0008662.
  4. Kamath R, Swain S, Pattanshetty S, Nair S. Studying risk factors associated with human Leptospirosis. Journal of Global Infectious Diseases. 2014;6(1):3-9.
  5. Haake DA, Levett PN. Leptospirosis in humans. Curr Top Microbiol Immunol. 2015;387:65–97.
  6. Vincent AT, Schiettekatte O, Goarant C, Neela VK, Bernet E, Thibeaux R, et al. Revisiting the taxonomy and evolution of pathogenicity of the genus Leptospira through the prism of genomics. PLoS Negl Trop Dis. 2019;13(5):e0007270.
  7. Uavechanichkul R, Sraphet S, Suwancharoen D, Triwitayakorn K. PCR-based technique for detection and differentiation of pathogenic and saprophytic Leptospira species. Int.J.Microbiol.Res. 2011;2(1):43-8.
  8. Karcher D, Grenfell RC, Moreno AM, Moreno LZ, Vasconcellos SA, Heinemann MB, et al. Identification of pathogenic and nonpathogenic Leptospira species of Brazilian isolates by matrix assisted laser desorption/ionization and time flight mass spectrometry. Brazilian journal of Microbiology. 2018;49(4):900–8.
  9. Lilenbaum W, Kremer F, Ristow P, Dellagostin O, Bourhy P, Hartskeerl R, et al. Molecular characterization of the first leptospires isolated from goats in Brazil. Braz J Microbiol. 2014;45(4):1527–30.
  10. Ferreira AS, Costa P, Rocha T, Amaro A, Vieira ML, Ahmed A, et al. Direct detection and differentiation of pathogenic Leptospira species using a multi-gene targeted real time PCR approach. PLOS ONE. 2014;9(11):e112312.
  11. Minister of Trade of the Republic of Indonesia. Regulation of the Minister of Trade of the Republic of Indonesia number : 70/M-DAG/PER/12/2013 Regarding Guidelines for the Arrangement and Development of Traditional Markets, Shopping Centers, and Modern Shops, Article 1. Jakarta: Ministry of Trade of the Republic of Indonesia; 2013.
  12. Sakundarno M, Bertolatti D, Maycock B, Spickett J, Dhaliwal S. Risk factors for Leptospirosis infection in humans and implications for public health intervention in Indonesia and the Asia-Pacific Region. Asia-Pacific J Public Heal. 2014;26(1):15-32.
  13. Monteiro MB, de Sousa IE, Piteira M, Coelho S, Freitas P. Leptospirosis, a re-emerging threat. Cureus. 2021;13(4): e14295.
  14. 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 Microbiology. 2020;20:59.
  15. Karuniawati A, Yasmon A, Ningsih I. Optimizing real-time PCR method to detect Leptospira spp. in human blood and urine specimens. Med J Indones. 2012;21(1):13-7.
  16. Ghazaei C. Pathogenic Leptospira: advances in understanding the molecular pathogenesis and virulence. Open Veterinary Journal. 2018;8(1): 13-24
  17. Hines MT. Leptospirosis in Chapter 32. Equine Infectious Diseases (Second Edition). 2014;302-311.e5.
  18. Andre-Fontaine G, Aviat F, Thorin C. Water borne leptospirosis: survival and preservation of the virulence of pathogenic Leptospira spp. in fresh water. CurrMicrobiol. 2015;71:136–42.
  19. Bierque E, Thibeaux R, Girault D, Soupe-Gilbert ME, Goarant C. A systematic review of Leptospira in water and soil environments. PLoS One. 2020;15(1):e0227055.
  20. Adler B. History of leptospirosis and leptospira. Curr Top Microbiol Immunol. 2015;387:1–9.

How to Cite

Hendrayana, M. A. (2023). Application of polymerase chain reaction (PCR) methods in distinguishing pathogenic and saprophytic Leptospira from the traditional market environment in Denpasar City. Indonesia Journal of Biomedical Science, 17(2), 135–140. https://doi.org/10.15562/ijbs.v17i2.473

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