Bahram Kazemi; Negar Seyed; Mojgan Bandehpour; Zarrin Sharifnia; Parviz Pakzad
Volume 5, Issue 3 , September 2008, , Pages 148-155
Abstract
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 ...
Read More
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 for use in an ELISA system designed to recognize the anti- C. trachomatis antibody in patient sera. Methods: The PCR product of the chlamydial omp2 gene was cloned in pBluescript and its first 1100 bp was sub-cloned in the pQE-30 expression vector and induced by IPTG. The recombinant protein was purified by affinity chromatography and its purity was confirmed by SDS-PAGE, gel diffusion and western blot analyses. The purified protein was coated onto a polysty-rene microplate and tested by ELISA using patient serum. Results: We have cloned, over-expressed and purified biologically functional recombinant truncated Omp2 from C. trachomatis for use, as a species-specific recognition antigen, in an ELISA system. In this study we determined a cut-off value of 0.345 for this ELISA system using 55 negative sera and measured six positive sera at dilutions of 1:20-1:2560. Conclusion: As a species-specific recognition antigen, the over-expressed and purified recombinant truncated Omp2 from C. trachomatis performed well in an ELISA system.
Habibollah Saadat; Parviz Pakzad; Mandana Sattari; Negar Seyed
Volume 1, Issue 1 , June 2004, , Pages 63-70
Abstract
Background: Streptokinase, which is injected intravenously with a standard dose of 1.5 MIU, is the most widely used thrombolytic agent around the world. What is so important about this bioproduct is the level of antistreptokinase (anti-sk) antibody in the population, which is directly correlated to the ...
Read More
Background: Streptokinase, which is injected intravenously with a standard dose of 1.5 MIU, is the most widely used thrombolytic agent around the world. What is so important about this bioproduct is the level of antistreptokinase (anti-sk) antibody in the population, which is directly correlated to the incidence of streptococcal infections in that population. Objective: Since Iran is an endemic area for streptococcal infections, this study was conducted to assess the anti-sk level in an Iranian population. Materials and Methods: 97 males and 47 females referred to Modarress Hospital of Tehran for coronary angiography and cardiac catheterization were included. 10 ml of venous blood was taken before angiographies from each patient. According to the angiography reports, the patients were divided into three groups: Coronary Artery Diseases (CAD, n=95), Rheumatic Heart Disease (RHD, n=19) and normal coronaries (n=30). The anti-sk antibody level was assessed in the serum samples of all patients using Enzyme Linked Immunosorbant Assay. Results: In 23.2% of patients with CAD, 40% of normal coronaries and 73.7% of patients with RHD, the serum samples contained more than 2 arbitrary units (AU) of anti-SK antibody which regarded as high levels. There was no significant difference between the anti-sk level of patients with CAD and normal coronaries (2.03 ± 3.02 AUs vs. 2.52 ± 2.23 AU), but the level of antibody in RHD group (8.16 ± 10.1 AU) was significantly higher than other groups (p<0.05). No significant correlation was observed between antibody levels and the age or gender of patients. Conclusion: We concluded that the level of anti-sk antibody is high in Iranian population as compared to other endemic areas for streptococcal infections. Also we found no relation between the level of antibody and sex and age of patients. This study accentuated the necessity of assessment of drug efficacy in endemic areas for streptococcal infections especially in those patients with valvular heart disease.