Leptospiral proteins expressed during acute & convalescent phases of human leptospirosis.
OBJECTIVE
The available serological techniques for the diagnosis of leptospirosis have less sensitivity during the early stage of the disease. Understanding of leptospiral proteins expressed during acute and convalescent phases of leptospirosis, would be help the develop of new serodiagnostic strategies. Therefore, the present study was carried out to identify (i) an antigen that is conserved among the various pathogenic leptospira; (ii) best protein antigen to which immune response can be identified in the acute phase; and (iii) best protein antigen which is present in convalescent sera which can be used for seroepidemiological studies.


METHODS
Quantitative immunoblot analysis was performed using acute and convalescent phase human sera along with sera from normal healthy individuals and from patients with typhoid, malaria and hepatitis as the controls. All the samples were analyzed for the leptospiral protein recognition by using IgM and IgG immunoblots. Leptospiral cell fractionation was performed using triton X-114 and lysozyme and further the conservation of leptospiral proteins was also performed.


RESULTS
In confirmed cases of leptospirosis, the IgG recognition in acute phase sera was 30.2, 39.5, 27.9, 55.8 and 27.9 per cent for the leptospiral proteins p32, p41/42, p58, p62 and p82 respectively. The IgG has considerably increased to 65.1, 55.8, 46.5, 67.4 and 48.8 per cent against the same proteins during convalescent phase. The IgM recognition was 32.6 , 32.6, 30.2 and 37.2 per cent for acute phase sera and 32.6, 37.2, 44.2 and 41.9 per cent for convalescent phase sera for the leptospiral proteins p14, p25, p32 and p41/42, respectively. Leptospiral proteins like p62 and p82 were recognized among all the control groups with 3.3-15.3 per cent for IgG recognition.


CONCLUSIONS
Leptospiral protein p32 was found to be highly sensitive and specific and could be useful for the development of newer techniques for diagnosis and seroepidemiological studies. Combination of p32 and p41/42 for IgG and p14, p25, p32, p41/42 for IgM would increase the sensitivity of these techniques further.
National Leptospirosis Reference Centre, Regional Medical Research Centre (ICMR)Port Blair, India.
K
K
Natarajaseenivasan
P
P
Vijayachari
A P
AP
Sugunan
S
S
Sharma
S C
SC
Sehgal
D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002458:Cell Fractionation; D002498:Centrifugation; D000090266:Chromobox Protein Homolog 5; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007919:Leptospira / Q000276:immunology* / Q000378:metabolism; D007922:Leptospirosis / Q000175:diagnosis* / Q000276:immunology / Q000378:metabolism*; D009113:Muramidase; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
D000942:Antigens, Bacterial; C000715594:CBX5 protein, human; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D009687:Nuclear Proteins; D000090266:Chromobox Protein Homolog 5; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D009113:Muramidase