Isolation by culture and PCR identification of LipL32 gene of pathogenic Leptospira spp. in wild rats of Kuala Lumpur

Malays J Pathol. 2017 Aug;39(2):161-166.

Abstract

Background: A study was conducted to confirm the status of rats as the carrier of pathogenic leptospira in Kuala Lumpur, Malaysia.

Method: A total of 140 urine samples were collected from trapped rats. These samples were cultured in EMJH enriched media and 18 of these samples (12.9%) were found to be positive when observed under x40 by dark field microscope. Genomic DNA was extracted from all the 18 native isolates for PCR.

Result: All the 18 isolates generated the expected 786 base pair band when the set of primers known to amplify LipL32 gene were utilized. These results showed that the primers were suitable to be used for the identification of pathogenic leptospira from the 18 rat samples.

Conclusion: The sequencing of the PCR products and BLAST analysis performed on each representative isolates confirmed the pathogenic status of all these native isolates as the LipL32 gene was detected in all the Leptospira isolates. This indicates that the rats are carriers of the pathogenic leptospira in the study area, and therefore are of public health importance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Wild / microbiology*
  • Bacterial Outer Membrane Proteins / genetics*
  • Bacterial Outer Membrane Proteins / isolation & purification
  • Disease Vectors*
  • Humans
  • Leptospira / genetics*
  • Leptospirosis / transmission*
  • Lipoproteins / genetics*
  • Lipoproteins / isolation & purification
  • Malaysia
  • Polymerase Chain Reaction
  • Rats / microbiology*

Substances

  • Bacterial Outer Membrane Proteins
  • LipL32 protein, Leptospira
  • Lipoproteins