BACKGROUND: Synthetic biology is emerging as a viable alternative for the production of recombinant antigens for diagnostic applications. It offers a safe alternative for the synthesis of antigenic principles derived from organisms that pose a high biological risk. METHODS: Here, we describe an enzyme-linked immunosorbent assay (ELISA) using the synthetic recombinant LipL32 (rLipL32) protein expressed in Escherichia coli for the detection of Leptospira-specific antibodies in human serum samples. The rLipL32-based ELISA was compared with a microscopic agglutination test (MAT), which is currently used as the gold standard for the diagnosis of leptospirosis. RESULTS: Our results showed that all the MAT-positive serum samples were positive for Leptospira-specific IgG in an ELISA, while 65% (n = 13) of these samples were also positive for Leptospira-specific IgM. In the MAT-negative serum samples, 80% and 55% of the samples were detected as negative by an ELISA for Leptospira-specific IgM and IgG, respectively. CONCLUSION: An ELISA using the synthetic rLipL32 antigen was able to distinguish Leptospira-specific IgM (sensitivity 65% and specificity 80%) and IgG (sensitivity 100% and specificity 55%) in human serum samples and has the potential to serve as a rapid diagnostic test for leptospirosis.
BACKGROUND: Synthetic biology is emerging as a viable alternative for the production of recombinant antigens for diagnostic applications. It offers a safe alternative for the synthesis of antigenic principles derived from organisms that pose a high biological risk. METHODS: Here, we describe an enzyme-linked immunosorbent assay (ELISA) using the synthetic recombinant LipL32 (rLipL32) protein expressed in Escherichia coli for the detection of Leptospira-specific antibodies in human serum samples. The rLipL32-based ELISA was compared with a microscopic agglutination test (MAT), which is currently used as the gold standard for the diagnosis of leptospirosis. RESULTS: Our results showed that all the MAT-positive serum samples were positive for Leptospira-specific IgG in an ELISA, while 65% (n = 13) of these samples were also positive for Leptospira-specific IgM. In the MAT-negative serum samples, 80% and 55% of the samples were detected as negative by an ELISA for Leptospira-specific IgM and IgG, respectively. CONCLUSION: An ELISA using the synthetic rLipL32 antigen was able to distinguish Leptospira-specific IgM (sensitivity 65% and specificity 80%) and IgG (sensitivity 100% and specificity 55%) in human serum samples and has the potential to serve as a rapid diagnostic test for leptospirosis.
Authors: Sohini Dey; C Madhan Mohan; T M A Senthil Kumar; P Ramadass; A Mahalinga Nainar; K Nachimuthu Journal: Vet Microbiol Date: 2004-10-05 Impact factor: 3.293
Authors: Cláudia P Hartleben; Fernanda M A Leal; Leonardo G Monte; Daiane D Hartwig; Fabiana K Seixas; Sílvio A Vasconcellos; Bibiana Brihuega; Odir A Dellagostin Journal: Curr Microbiol Date: 2012-10-14 Impact factor: 2.188
Authors: Daiane D Hartwig; Thaís L Oliveira; Fabiana K Seixas; Karine M Forster; Caroline Rizzi; Cláudia P Hartleben; Alan J A McBride; Odir A Dellagostin Journal: Microb Cell Fact Date: 2010-12-06 Impact factor: 5.328
Authors: Alan Villalobos; Jon E Ness; Claes Gustafsson; Jeremy Minshull; Sridhar Govindarajan Journal: BMC Bioinformatics Date: 2006-06-06 Impact factor: 3.169