title: Surface-associated Hsp60 chaperonin of Leptospira interrogans serovar Autumnalis N2 strain as an immunoreactive protein.
abstract: The present study has been formulated in order to detect an immunoreactive protein whose identification can play a major role in the early diagnosis of disease. The identified protein will be produced by recombinant methods and used for the recombinant protein based ELISA. A comparison was made between the developed method and the gold standard MAT test to evaluate the serodiagnosis potential of the protein. The protein profile, immunoblot and MALDI-TOF analysis was carried out to identify the immunoreactive protein. The immunoreactive protein identified was used to develop ELISA for the diagnosis of leptospirosis using patients' sera with various clinical manifestations. The immunoreactive protein was identified as Leptospira GroEL chaperonin of molecular weight 60 kDa. The theoretical/experimental molecular weights, pI were found to be 58.5/60 kDa and 5.41/6, respectively. The overall results of the recombinant GroEL-IgM ELISAs showed cumulative sensitivity, specificity, positive predictive value, and negative predictive values of 90.6%, 94.9%, 94.6%, and 91.0%, respectively. The performance of such ELISA appeared better than that of any other serological tests previously evaluated for the diagnosis of leptospirosis in India. Thus, a highly conserved and immunogenic outer exposed GroEL protein during infection clearly merits further use in the serodiagnosis of leptospirosis.
authors.affiliation: Medical Microbiology Laboratory, Department of Microbiology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620 024, India. natarajaseenivasan@rediffmail.com
authors.forename: K
authors.initials: K
authors.lastname: Natarajaseenivasan
authors.forename: S C
authors.initials: SC
authors.lastname: Artiushin
authors.forename: S
authors.initials: S
authors.lastname: Velineni
authors.forename: K
authors.initials: K
authors.lastname: Vedhagiri
authors.forename: P
authors.initials: P
authors.lastname: Vijayachari
authors.forename: J F
authors.initials: JF
authors.lastname: Timoney
mesh_terms: D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D002457:Cell Extracts / Q000276:immunology; D018834:Chaperonin 60 / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D041301:Leptospira interrogans serovar autumnalis / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification; D007922:Leptospirosis / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002457:Cell Extracts; D018834:Chaperonin 60; D011994:Recombinant Proteins