Leptospiral phosphatidylinositol-specific phospholipase C: a novel conserved antigen for the definitive diagnosis of Leptospirosis.

Abstract

Identification and characterisation of novel antigenic proteins of Leptospira are necessary to unravel the secrets of the Leptospiral disease biology. Besides, the current scenario of Leptospirosis demands the development of advanced diagnostics with the potential of an early and specific diagnosis. This work reports the diagnostic efficacy of a PI-PLC (LA_1375) of Leptospira interrogans serovar Lai strain 56601. Phospholipases are enzymes that catalyse the degradation of membrane phospholipids, with the after effects of pathophysiological consequences through the impairment of cellular signalling mechanisms. The immunoblotting analysis allows to identify the Leptospirosis positive serum samples demonstrated the expression and exposure of the protein to the immune system and confirmed the antigenic nature of the protein. Furthermore, evaluation of diagnostic efficacy of an ELISA to detect antibodies against recombinant PI-PLC has tested in indirect IgM ELISA using the purified 50kDa recombinant protein as the coating antigen. A cut-off value of 0.218at mean + 2SD and a sensitivity value of 100% and a specificity value of 95.65% explored for the rPI-PLC IgM ELISA. The PPV and NPV of ELISA for the Leptospirosis confirmed acute phase samples were 95.87% and 100% respectively. Comparison of the ELISA assay with MAT for the detection of Leptospiral antibodies showed a positive correlation (r value 0.345, p < 0.05), and signified the diagnostic efficacy of rPI-PLC for the definitive diagnosis of the Leptospirosis. In addition, the revealed conservation nature of PI-PLC among different Leptospira species highlights the diagnostic and vaccine candidate potential. Thus, the study emphasises the Leptospiral PI-PLC as a potential antigen still the importance as a novel biomarker candidate for the serodiagnosis and pathogenesis of Leptospirosis have to be investigated.