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Protocol Name
DOI:DOI:10.17504/protocols.io.xdwfi7e RRID Copied  
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Qiang Wang, Yan Lei, Xiaolan Lu, Guangrong Wang, Qin Du, Xiaolan Guo, Quming Fan, Guoyuan Zhang, Dongsheng Wang 2019. Urea-mediated dissociation alleviate the false-positive Treponema pallidum-specific antibodies detected by ELISA. protocols.io https://dx.doi.org/10.17504/protocols.io.xdwfi7e
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Protocol Information

URL: https://dx.doi.org/DOI:10.17504/protocols.io.xdwfi7e

Authors: Qiang Wang, Yan Lei, Xiaolan Lu, Guangrong Wang, Qin Du, Xiaolan Guo, Quming Fan, Guoyuan Zhang, Dongsheng Wang

Summary: The serological detection of antibodies to Treponema pallidum is essential to the diagnosis of syphilis. However, for the presence of cross-reaction, the specific antibody tests [e.g., enzyme-linked immunosorbent assay (ELISA)] always have false-positive results. In this study, we derived and validated the dissociation of urea in an attempt to alleviate the situation of false-positive antibodies to T. pallidum detected by ELISA. Six serum samples that were false-positive antibodies to T. pallidumdetected by ELISA, and 16 control serum samples (8 sera positive for both specific IgG and IgM, and 8 IgG-positive and IgM-negative sera) were collected to select the appropriate dissociated concentration and time of urea. Our goal was to establish improved an ELISA method based on the original detection system of ELISA. The sensitivity of the improved ELISA was evaluated by 275 serum samples with class IgM-positive antibodies to T. pallidum. At 6 mol/L with 10 minutes dissociation of urea, 6 samples with false-positive antibodies to T. pallidum were converted to negative, and compared with true-positive antibodies to T. pallidum. The sensitivity of the improved ELISA was 100% by detecting the class IgM-positive antibodies to T. pallidum in sera of patients with syphilis. Considering the importance at the diagnosis of syphilis, antibodies to T. pallidumin serum samples should be retested by the improved ELISA method to avoid false-positive results.

Affiliations: Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College; Faculty of Laboratory Medicine, Center for Translational Medicine, North Sichuan Medical college, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College; Faculty of Laboratory Medicine, Center for Translational Medicine, North Sichuan Medical college, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College; Faculty of Laboratory Medicine, Center for Translational Medicine, North Sichuan Medical college, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College; Faculty of Laboratory Medicine, Center for Translational Medicine, North Sichuan Medical college, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College, Department of Laboratory Medicine, Affiliated Hospital of North Sichuan Medical College

Version: 1

Publication Date: 2019

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