An Insight for the Future Development of Diagnostic Tool by Exploiting Novel Leishmania Donovani Recombinant Hypothetical Protein
Journal Title: Journal of Clinical and Diagnostic Research - Year 2018, Vol 12, Issue 11
Abstract
ABSTRACT Introduction: Assenting diagnosis of Visceral Leishmaniasis (VL) relies on the detection of Leishmania donovani (L. donovani) in splenic and bone marrow specimens obtained by invasive techniques. Thus, the development of inexpensive, non-invasive serological test encompassing high specificity, sensitivity and diagnostic efficacy is urgently needed. Aim: To assess the significance of recombinant proteins possessing B-cell epitopes in VL diagnosis. Materials and Methods: Employing immunoinformatics approach, the B-cell epitope footprint of L. donovani hypothetical proteins (derived from earlier studies) were decrypted holding good antigenic character with numerous epitopes. L. donovani hypothetical proteins XP_003860226.1 and XP_003861271.1 were first time cloned as His-tagged fusion proteins and purified as novel recombinant protein antigens, designated rLdhyb and rLdhyc respectively. Sanger sequencing method was exploited to sequence gene insert (GeneBank accession number MH479406). Results: B-cell epitopes revealed 100% conservancy with L. infantum. Immunoinformatics data revealed no significant sequence similarity with homo sapien and the causative agent of other diseases like tuberculosis, typhoid, malaria etc., resembling in symptoms to VL. Sequencing chromatogram of cloned gene Ldhyb and Ldhyc revealed 98% and 94% identity with L. donovani. ELISA revealed the absolute specificity with sensitivity of 95.4% for rLdhyb and 91% for rLdhyc. Area under curve for rLdhyb, rLdhyc and SLA were 0.99, 0.99 and 0.961, with standard error 0.002, 0.007 and 0.019 respectively. The in silico data was coherently supported by in vitro result. Conclusion: Absolute specificity, high sensitivity and diagnostic efficacy (rLdhyb: 98%; rLdhyc: 97%) advocated their excellent biomarker property. The present findings provide some basic insights for the future development of novel hypothetical proteins based non-invasive diagnostic tool for VL detection.
Authors and Affiliations
Fauzia Jamal, Manish Kumar Singh, Jagadish Hansa, Pushpanjali . , Pushkar Shivam, Sarita Kumari, Shyam Narayan, Shubhankar K Singh
Practice of Intravenous to Oral Conversion of Antibiotics and its Influence on Length of Stay at a Tertiary Care Hospital: A Prospective Study
ABSTRACT Introduction: Several studies have demonstrated the efficacy and safety of switching from intravenous to oral antibiotics in clinically stable patients. Early switch from Intravenous (IV) to Per Oral (PO) could...
Evaluation of Efficacy of 1% Curcuminoids as Local Application in Management of Oral Lichen Planus – Interventional Study
Introduction: Oral Lichen Planus (OLP) is a chronic inflammatory mucocutaneous disease affecting the oral cavity which has been associated with severe burning sensation which in turn affects the quality of life of the pa...
Effect of Ultrasonic Bath Immersion on Physical Properties of an MMA-based Orthodontic Base-plate Material
ABSTRACT Introduction: Self-cured Methyl Methacrylate (MMA)-based acrylic resin is commonly used for fabricating removable orthodontic appliances. However, residual monomer can leach out and cause mucosal irritation. Fur...
Effect of Motives for Food Choice on Oral Health among Primary School Children in Mangalore: An Analytical Survey
Introduction: Parents influence children’s eating behaviours by making some foods available than others and by acting as models of eating behaviour. Food selected by parents influence general and oral health of their chi...
Deep Cerebral Vein Thrombosis: A Clinical Masquerader
Cerebral Vein Thrombosis (CVT) is an uncommon cause of stroke. Thrombosis can occur in superficial veins, deep venous system or cortical veins of brain. The term Deep Cerebral Vein Thrombosis (DCVT) is used for thrombosi...