Yaakob Yuszniahyati, Rodrigues Kenneth Francis, Opook Fernandes, William Timothy, John Daisy Vanitha
Biotechnology Research Institute, Universiti Malaysia Sabah, Kota Kinabalu, Sabah, Malaysia.
Jesselton Medical Centre, Kota Kinabalu, Sabah, Malaysia.
Malays J Med Sci. 2017 Oct;24(5):44-51. doi: 10.21315/mjms2017.24.5.5. Epub 2017 Oct 26.
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.
Here, we describe an enzyme-linked immunosorbent assay (ELISA) using the synthetic recombinant LipL32 (rLipL32) protein expressed in 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.
Our results showed that all the MAT-positive serum samples were positive for Leptospira-specific IgG in an ELISA, while 65% ( = 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.
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.
合成生物学正成为生产用于诊断应用的重组抗原的一种可行替代方法。它为合成源自具有高生物风险的生物体的抗原成分提供了一种安全的替代方案。
在此，我们描述了一种酶联免疫吸附测定（ELISA），该方法使用在 中表达的合成重组LipL32（rLipL32）蛋白来检测人血清样本中的钩端螺旋体特异性抗体。将基于rLipL32的ELISA与显微镜凝集试验（MAT）进行比较，MAT目前用作钩端螺旋体病诊断的金标准。
我们的结果表明，所有MAT阳性血清样本在ELISA中钩端螺旋体特异性IgG均为阳性，而这些样本中有65%（ = 13）的钩端螺旋体特异性IgM也为阳性。在MAT阴性血清样本中，分别有80%和55%的样本在ELISA中检测钩端螺旋体特异性IgM和IgG为阴性。
使用合成rLipL32抗原的ELISA能够区分人血清样本中的钩端螺旋体特异性IgM（敏感性65%，特异性80%）和IgG（敏感性100%，特异性55%），并且有潜力作为钩端螺旋体病的快速诊断检测方法。
Zotero 插件