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IDENTIFICATION OF L. DONOVANI SPECIFIC 70 KDA, 37 KDA AND 12.6 KDA PROMASTIGOTE ANTIGENS FOR THE DIAGNOSIS OF INDIAN VISCERAL LEISHMANIASIS'.
IDENTIFICATION OF L. DONOVANI SPECIFIC 70 KDA, 37 KDA AND 12.6 KDA PROMASTIGOTE ANTIGENS FOR THE DIAGNOSIS OF INDIAN VISCERAL LEISHMANIASIS'.
Diagnosis of Visceral Leishmaniasis (VL) is based on microscopic demonstration of amastigote in splenic/bone marrow smears, but these are painful and risky process. Serodiagnosis using recombinant k39 antigen has been a significant advance in management of VL, however, being positive in 20-32% healthy individuals is an important drawback. In this study we have identified three novel antigens of Leishmania donovani comprising of 70 kDa, 37 kDa and 12 kDa protein through western blotting. We analyzed the antibody response against crude soluble antigen (CSA) of L. donovani in VL patients (before and after treatment), in endemic healthy (EHC) and non endemic healthy controls (NEHC). Our results suggest that the antibody response against this antigen is predominantly found in serum of all VL patients even after the successful treatment but is not present in the endemic and non endemic healthy controls. To validate immunoblotting results, we partially purified this antigen by gel elution and performed indirect enzyme linked immunosorbent assay (ELISA) on VL patients, NEHC, EHC and different disease controls. The sensitivity and specificity of all three proteins in case of viceral leishmaniasis, endemic healthy, non endemic healthy and different disease group was described above respectively. We further characterized these proteins through 2D gel electrophoresis and MALDI-TOF followed by trypsin digestion. Results suggest that the BHU P-l protein is a heat shock protein of 70 kDa and 653 amino acids, second is BHU P-2 protein of 37 kDa which is new one in case leishmania donovani and third one is of 12 kDa and 113 amino acids is also a new protein in case of leishmania donovani. Because of their high sensitivity and specificity, these antigens have great potential for further development as tool for diagnosis of VL.
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