Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples
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 enzy...
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Universiti Sains Malaysia
2017
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Online Access: | https://eprints.ums.edu.my/id/eprint/30245/1/Recombinant%20LipL32%20protein%20developed%20using%20a%20synthetic%20gene%20detects%20leptospira-specific%20antibodies%20in%20human%20serum%20samples%20ABSTRACT.pdf https://eprints.ums.edu.my/id/eprint/30245/2/Recombinant%20LipL32%20protein%20developed%20using%20a%20synthetic%20gene%20detects%20leptospira-specific%20antibodies%20in%20human%20serum%20samples%20FULL%20TEXT.pdf https://eprints.ums.edu.my/id/eprint/30245/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5772814/pdf/05mjms24052017_oa3.pdf https://doi.org/10.21315/mjms2017.24.5.5 |
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my.ums.eprints.302452021-08-21T04:09:34Z https://eprints.ums.edu.my/id/eprint/30245/ Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples Yuszniahyati Yaakob Kenneth Francis Rodrigues Fernandes Opook Timothy William Daisy Vanitha John RB37-56.5 Clinical pathology. Laboratory technique 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. Universiti Sains Malaysia 2017 Article PeerReviewed text en https://eprints.ums.edu.my/id/eprint/30245/1/Recombinant%20LipL32%20protein%20developed%20using%20a%20synthetic%20gene%20detects%20leptospira-specific%20antibodies%20in%20human%20serum%20samples%20ABSTRACT.pdf text en https://eprints.ums.edu.my/id/eprint/30245/2/Recombinant%20LipL32%20protein%20developed%20using%20a%20synthetic%20gene%20detects%20leptospira-specific%20antibodies%20in%20human%20serum%20samples%20FULL%20TEXT.pdf Yuszniahyati Yaakob and Kenneth Francis Rodrigues and Fernandes Opook and Timothy William and Daisy Vanitha John (2017) Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples. Malaysian Journal of Medical Science, 25. pp. 44-51. ISSN 1394-195X (P-ISSN) , 2180-4303 (E-ISSN) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5772814/pdf/05mjms24052017_oa3.pdf https://doi.org/10.21315/mjms2017.24.5.5 |
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RB37-56.5 Clinical pathology. Laboratory technique Yuszniahyati Yaakob Kenneth Francis Rodrigues Fernandes Opook Timothy William Daisy Vanitha John Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
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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. |
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Article |
author |
Yuszniahyati Yaakob Kenneth Francis Rodrigues Fernandes Opook Timothy William Daisy Vanitha John |
author_facet |
Yuszniahyati Yaakob Kenneth Francis Rodrigues Fernandes Opook Timothy William Daisy Vanitha John |
author_sort |
Yuszniahyati Yaakob |
title |
Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
title_short |
Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
title_full |
Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
title_fullStr |
Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
title_full_unstemmed |
Recombinant LipL32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
title_sort |
recombinant lipl32 protein developed using a synthetic gene detects leptospira-specific antibodies in human serum samples |
publisher |
Universiti Sains Malaysia |
publishDate |
2017 |
url |
https://eprints.ums.edu.my/id/eprint/30245/1/Recombinant%20LipL32%20protein%20developed%20using%20a%20synthetic%20gene%20detects%20leptospira-specific%20antibodies%20in%20human%20serum%20samples%20ABSTRACT.pdf https://eprints.ums.edu.my/id/eprint/30245/2/Recombinant%20LipL32%20protein%20developed%20using%20a%20synthetic%20gene%20detects%20leptospira-specific%20antibodies%20in%20human%20serum%20samples%20FULL%20TEXT.pdf https://eprints.ums.edu.my/id/eprint/30245/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5772814/pdf/05mjms24052017_oa3.pdf https://doi.org/10.21315/mjms2017.24.5.5 |
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1760230737795088384 |
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13.18916 |