Molecular Detection of Leptospira Using Polymerase Chain Reaction Based on LipL32 and rpoB Gene

Chandan, S. and Umesha, S. (2014) Molecular Detection of Leptospira Using Polymerase Chain Reaction Based on LipL32 and rpoB Gene. Research and Reviews: Journal of Veterinary Science and Technology, 3 (1). pp. 16-23.

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Abstract

Leptospirosis is an emerging disease for which culture and identification are partly unresolved. In fact, 16S rRNA-based sequencing is the most widely used PCR methodology that can detect such uncultivable pathogens. However, this assay has some limitations linked to potential problems of contamination, which hampers diagnosis. To overcome this, the authors have a simple PCR strategy involving targeting of the gene encoding the LipL32 and RNA polymerase β subunit (rpoB), a highly conserved enzyme. We report here a case study of Leptospira-infected samples; Leptospira has been isolated from biological samples. Leptospirosis is a septicemia bacterial disease affecting animals and human being, the infection may be symptomatic, mild or severe and acute or chronic. PCR assay targeting partial rpoB and LipL32 gene of Leptospira using primers, with a product size of 600 and 532 base pair long which are Leptospira specific, where rpoB detects both pathogenic and non-pathogenic and efficacy of LipL32 primer capable of amplifying conserved outer membrane protein (OMP) gene. LipL32 is the major leptospiral outer membrane lipoprotein expressed during infection. The sequence of the Leptospira rpoB gene was determined and compared with the published sequence. The findings have significant implications for the development of a new tool for identification of spirochetes, especially if clinical samples are contaminated.

Item Type: Article
Subjects: Agriculture Biological Sciences > Biotechnology
University of Mysore > PG Campuses > Manasagangotri, Mysore > Agriculture Biological Sciences > Biotechnology
Divisions: PG Campuses > Manasagangotri, Mysore > Biotechnology
Depositing User: Praveen Kumari B.L
Date Deposited: 06 Feb 2015 06:54
Last Modified: 06 Feb 2015 06:54
URI: http://eprints.uni-mysore.ac.in/id/eprint/17503

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