A sensitive and specific IgM-ELISA for the serological diagnosis of human leptospirosis using a rLipL32/1-LipL21-OmpL1/2 fusion protein.
- Author:
Aihua SUN
1
;
Yuan WANG
;
Peng DU
;
Shengling WU
;
Jie YAN
Author Information
- Publication Type:Journal Article
- MeSH: Antigens, Bacterial; genetics; metabolism; Bacterial Outer Membrane Proteins; genetics; metabolism; Enzyme-Linked Immunosorbent Assay; methods; Humans; Immunoglobulin M; immunology; Leptospirosis; diagnosis; metabolism; Lipoproteins; genetics; metabolism; Recombinant Fusion Proteins; genetics; metabolism
- From: Biomedical and Environmental Sciences 2011;24(3):291-299
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo construct a lipL32/1-lipL21-OmpL1/2 fusion gene and its prokaryotic expression system, and to establish an enzyme-linked immunosorbent assay (ELISA) using the rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for sensitive and specific detection of IgM in the serum of patients with leptospirosis.
METHODSlipL32/1-lipL21-OmpL1/2 fusion genes were constructed using a primer-linking PCR. The target recombinant protein antigens, rLipL32/1, rLipL21, rOmpL1/2 and rLipL32/1-LipL21-OmpL1/2, were expressed and the purified antigens were then immobilized to the surface of microplate wells for ELISA-based detection of IgM in the sera of leptospirosis patients.
RESULTSOf 493 acute leptospirosis patients, 95.7% and 97.8% were positive by rLipL32/1-LipL21- OmpL1/2-IgM-ELISA using different serum dilutions, which was higher than the rLipL32/1-IgM-ELISA (93.1% and 90.3%), rLipL21-IgM-ELISA (90.3% and 87.0%), and rOmpL1-IgM-ELISA (85.6% and 81.1%) (P<0.01). All IgM-ELISAs tested negative against 56 non-leptospirosis patients with typhoid fever, hemorrhagic fever or dengue fever.
CONCLUSIONTrigeminal fusion antigen increases ELISA sensitivity and the rLipL32/1-LipL21-OmpL1/2- IgM-ELISA is a sensitive and specific serological diagnostic method for clinical leptospirosis.
