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Sequence analysis of LipL41 and LipL21: Prospective Outer Membrane Proteins (OMPs) in early diagnosing leptospirosis
Muzaffar Mosquill1, Syafinaz Amin Nordin1, Mohamad Ridhuan Mohd Ali2
1Department of Medical Microbiology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Selangor 43400, Malaysia.
Abstract:
Leptospirosis is a zoonotic disease mostly occurring in tropical climate countries. The etiology of the disease is due to microbes from the genus Leptospira. Higher number of cases reported worldwide indicated the disease is not easily eradicated. Leptospirosis shares the most common febrile symptoms such as dengue, Zika and yellow fever thus making it difficult to differentiate the disease at an early stage. The widely used current detection via PCR, uses the bacterial outer membrane protein (OMP) as their target region. However, the heterogeneity and variation of the genome cause false negative results. Lipoprotein LipL41 is the third most abundant outer membrane lipoprotein among pathogenic species and it is surface exposed and expressed during infection thus making it a suitable candidate in identifying pathogenic Leptospira. LipL21 on the other hand is a potential candidate in identifying the intermediate species. The study aimed in designing suitable PCR primers in identifying pathogenic and intermediate species of Leptospira through bioinformatics analysis on the bacterial OMPs. LipL41 and LipL21 were chosen as the suitable target sequence to be used as PCR primers in detecting the pathogenic and intermediate species, respectively. The designed primers indicated positive feedback upon tested with their respective bacterial DNA extract. These lipoproteins may serve as potential PCR primers to be used with clinical samples in diagnosing leptospirosis.•The etiology of the illness is due to bacteria from the genus Leptospira.•PCR utilizes the bacterial external membrane protein (OMP) thus the heterogeneity and variety of the genome cause bogus adverse outcomes.•The suitable candidates are LipL41, the third most abundant outer membrane lipoprotein, whereas LipL21 is a potential candidate in identifying the intermediate species.
Insights
New PCR primers targeting LipL41 and LipL21 lipoproteins can accurately detect pathogenic and intermediate Leptospira species. This improves leptospirosis diagnosis, overcoming limitations of current methods that suffer from false negatives due to genomic variations.
Area of Science:
- Microbiology
- Infectious Diseases
- Bioinformatics
Background:
- Leptospirosis is a significant zoonotic disease caused by *Leptospira* bacteria, prevalent in tropical regions.
- Current diagnostic methods like Polymerase Chain Reaction (PCR) targeting outer membrane proteins (OMPs) can yield false negatives due to bacterial genomic heterogeneity.
- Accurate early diagnosis is challenging as leptospirosis shares symptoms with other febrile illnesses such as dengue and Zika.
Purpose of the Study:
- To design novel PCR primers for identifying pathogenic and intermediate *Leptospira* species.
- To utilize bioinformatics analysis of bacterial OMPs for primer design.
- To address the limitations of existing diagnostic techniques for leptospirosis.
Main Methods:
- Bioinformatic analysis was performed on bacterial OMPs of *Leptospira*.
- Lipoprotein LipL41 was selected as a target for pathogenic species, and LipL21 for intermediate species.
- PCR primers were designed based on these selected target sequences.
Main Results:
- The designed primers demonstrated positive results when tested against their respective bacterial DNA extracts.
- LipL41 and LipL21 proved effective as targets for detecting pathogenic and intermediate *Leptospira* species, respectively.
- The developed primers show potential for improved diagnostic accuracy in clinical samples.
Conclusions:
- LipL41 and LipL21 are suitable targets for developing specific PCR primers for *Leptospira* detection.
- These novel primers offer a promising alternative for accurate diagnosis of leptospirosis.
- The study highlights the potential of these lipoproteins for use in clinical diagnostic assays.
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