Antigen capture ELISA for the heat shock protein (hsp60) of Chlamydia trachomatis.
نویسندگان
چکیده
AIMS To develop an indirect ELISA using the heat shock protein (hsp60) of Chlamydia trachomatis as antigen. METHODS The hsp60 gene was amplified by PCR, expressed in the vector pDEV-107 and transformed into Escherichia coli. The recombinant protein, expressed as a beta-galactosidase fusion product, was captured onto a solid phase using a monoclonal antibody directed against beta-galactosidase. Following incubation with goat anti-human antibody conjugated to peroxidase and colour development on addition of peroxidase substrate, antibody recognition of antigen was quantified by optical density at 492 nm. RESULTS A sensitive and relatively specific ELISA to detect hsp60 has been produced, which can be exploited to determine the antibody response to C trachomatis hsp60. CONCLUSIONS This assay will permit the future investigation of the immunopathogenesis of persistent inflammation following C trachomatis infection.
منابع مشابه
High Sensitivity C–Reactive Protein and Im-munoglobulin G against Chlamydia Pneumo-niae and Chlamydial Heat Shock Protein-60 in Ischemic Heart Disease
Background: Inflammation and infectious agents such as Chlamydia pneumoniae have been associated with cardiovascular disease. Objective: To evaluate the serum high sensitivity C - reactive protein (hs-CRP) and antibodies against Chlamydia pneumoniae and Chlamydial heat shock protein-60 (Cp-HSP60) in patients with ischemic heart disease (IHD). Methods: 62 patients with IHD having either acute my...
متن کاملHumoral immune response to conserved epitopes of Chlamydia trachomatis and human 60-kDa heat-shock protein in women with pelvic inflammatory disease.
The association between humoral immunity to unique and conserved epitopes of the Chlamydia trachomatis 60-kDa heat-shock protein (hsp60) and immunity to human hsp60 was examined in 129 women with laparoscopically verified pelvic inflammatory disease. An ELISA was used to detect antichlamydial IgG and IgA antibodies, IgG antibodies to recombinant human hsp60, and antibodies to two synthetic pept...
متن کاملContinuous B-cell epitopes in Chlamydia trachomatis heat shock protein 60.
B-cell peptide epitopes in chlamydial heat shock protein 60 (hsp60) were elucidated with antisera from 13 rabbits immunized with Chlamydia trachomatis serovars B, C, and L2 and antisera from eight women with C. trachomatis-associated ectopic pregnancies. Thirteen major epitopes were identified with the human sera, 10 of which were also observed with rabbit antisera. Seven of the 13 epitopes rec...
متن کاملAutoimmunity to heat shock protein 60 and antigen-specific production of interleukin-10.
The immunopathologic sequelae of chlamydial infection are correlated with immune responses to the Chlamydia trachomatis heat shock protein 60 (hsp60). One pathogenic mechanism that may explain this association is the induction of autoimmune responses to self hsp60, since these two proteins share a high degree of amino acid sequence identity. To investigate the conditions under which autoimmune ...
متن کاملCloning, Expression and Purification of Truncated Chlamydia Trachomatis Outer Membrane Protein 2 (Omp2) and its Application in an ELISA Assay
Background: Although a simple and direct method does not exist for the detection of chlamydial infections, there are situations in which reliable serological tests, with sensi-tivity related to a specific antigen, could be helpful. Objective: The aim of this study was to clone the first 1100 bp of the C. trachomatis outer membrane protein 2 (omp2) gene in order to prepare a recombinant protein ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of clinical pathology
دوره 49 8 شماره
صفحات -
تاریخ انتشار 1996