International Journal of Pharma and Bio Sciences
 
 
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ORIGINAL RESEARCH ARTICLE
Int J Pharm Bio Sci Volume 12 Issue 3, 2021 (July-September), Pages:86-93

Molecular and Immunological Surveillance of Leptospira Incidence at Gwalior Region of Central India

Bharti Mankere, Preeti Gupta, Kundan Tandel, Urmil Tuteja , Mahendra K Gupta and Kulanthaivel Thavachelvum
DOI: http://dx.doi.org/10.22376/ijpbs.2021.12.3.B86-B93
Abstract:

Leptospirosis is an infection caused by the spirochete bacteria Leptospira.  It is ubiquitous around the world and in India it has been reported from several parts of the country during the rainfall season. In our present work, a detailed cross sectional study with the serum samples from pyrexia of unknown origin (PUO) patients around the poorly studied Gwalior district of the central India region was undertaken to assess the Leptospirosis incidence. Immunological methods such as MAT (Microscopic agglutination test), ELISA (Enzyme linked immune sorbent assay), and WB (Western blotting), along with molecular methods such as PCR and 2D protein analysis were employed and it was found that about 2.82% (n=854) of samples tested were positive for Leptospirosis. The antibody response was obtained against Leptospira interrogans serovars Bataviae, Ballum, Cyanoptera, Grippotyphosa and Ictero-haemorrhagiae copenhageni. The 2-D Gel electrophoresis of the serovar Copenhageni when probed with positive sera in WB identified a major antigen that provoked the immune response in patients. The reaction corresponding to the 32-kDa size protein and an isoelectric point pI value of 6.5 indicated that the immune response was against the outer membrane protein lipL32 of Leptospira spp. Further confirmation with the MALDI TOF analysis of the excised sample from the parallel gel confirmed the tryptic digested products of lipL32. Isolation of leptospires could not be successful; however, the detection of leptospiral DNA by PCR using lipL32 primers in the MAT positive samples confirmed the etiology of the disease. The present study suggests that Leptospirosis could be considered as one of the prime suspects, in the PUO cases, around Gwalior region of Central India.

Keywords: Leptospirosis, PUO cases, MAT, PCR, MALDI –TOF, LipL32.
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