Diagnosis of leptospirosis by recombinant antigen based single serum dilution ELISA.
OBJECTIVE
Leptospirosis, a zoonosis with a worldwide distribution is an acute febrile illness caused by spirochaetes of the pathogenic Leptospira interrogans. Microscopic agglutination test (MAT), the reference method for diagnosis was successively done to evaluate the modified ELISA which was developed with the recombinant LipL32 antigen for the detection of anti-leptospiral antibodies in human serum samples.


METHODS
The recombinant LipL32 antigen was developed from the serovar Pomona strain Pomona of the pathogenic L. interrogans species. The predicted titre at a single working dilution was plotted against the observed antiserum titre. Subsequently, predicted antibody activity titres were determined directly from the standard curve by solving the regression line equation. The relative sensitivity, specificity and accuracy of the single dilution ELISA for the detection of anti-leptospiral antibodies were determined in comparison to the MAT.


RESULTS
A linear relationship was found between the predicted antibody titres at a single working dilution of 1:250 and the corresponding observed serum titres by the standard serial-dilution method. Regression analysis was used to determine a standard curve from which an equation was derived that allowed demonstration of the mentioned correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. A high level of sensitivity of 96 per cent and specificity of 91 per cent between ELISA and MAT titres was found. The kappa value was almost 1.0 indicating perfect agreement.


CONCLUSIONS
The r LipL32 ELISA was proved to be sensitive, specific and accurate as compared to the standard MAT and the test could be efficiently utilized as a screening test for a large number of human serum samples for the detection of leptospiral antibodies.
Dey
Sohini
S
Division of Animal Biotechnology, Indian Veterinary Research Institute, Bareilly, India. sohinimadhan@yahoo.com
Mohan
C Madhan
CM
Ramadass
P
P
Nachimuthu
K
K
D000942:Antigens, Bacterial / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007921:Leptospira interrogans / Q000276:immunology*; D007922:Leptospirosis / Q000175:diagnosis*; D008074:Lipoproteins / Q000097:blood; D012044:Regression Analysis; D012680:Sensitivity and Specificity; D015927:Skin Test End-Point Titration
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C406589:LipL32 protein, Leptospira; D008074:Lipoproteins