Detection of Leptospira species targeting 16S rRNA, rpoB, and lipL32 gene from clinical and environmental sources
Leptospirosis is currently emerging globally as reflected by the increase in the number of Leptospira infections. The scope of research on identification of individual Leptospira species through a rapid assay in local setting is still lacking. This study attempts to determine the Leptospira speci...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2019
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/84213/1/FPSK%20%28m%29%202019%2038%20UPM%20ir.pdf |
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| Summary: | Leptospirosis is currently emerging globally as reflected by the increase in the
number of Leptospira infections. The scope of research on identification of
individual Leptospira species through a rapid assay in local setting is still lacking.
This study attempts to determine the Leptospira species distribution in clinical
and environmental samples using molecular approach. The blood for clinical
samples (n=64) were collected from patients whom have been suspected with
leptospirosis, and the environmental samples (n=105) consisting of soils and
waters were collected from areas with potential sources of Leptospira
transmission. All samples were directly extracted for DNA and subjected to PCR
using three sets of established primers; 16S rRNA, rpoB, and lipL32. Positive
samples were further analysed on their DNA sequences and also through
phylogenetic analysis. Six clinical samples were amplified for 16S rRNA gene
and four clinical samples showed an amplification of rpoB gene that was highly
matched to pathogenic Leptospira spp. in GenBank. Out of 13 positive results
amplified for 16S rRNA from environmental samples, only five were detected to
have been contaminated by Leptospira spp. while the others showed a higher
number of contamination by different species of bacteria. The BLAST similarity
results for clinical samples targeting lipL32 showed that only one out of 64
samples was highly matched to pathogenic Leptospira spp. in GenBank. The
phylogenetic trees constructed shows that the positive clinical samples were
clustered into group with pathogenic Leptospira spp. while the environmental
samples showed a clustering into uncultured Leptospira. Overall, this study
showed the ability of the molecular approach in combination with PCR in
determine Leptospira species directly on both clinical and environmental
samples. Although the number of positive samples were low, but the tendency
of the species to appear in limited positive samples may infer the species as the
likely agents in causing leptospirosis at this study setting. |
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