Reliable tool for detection of novel Coxiella burnetii antigens, using immobilized human polyclonal antibodies.

Coxiella burnetii (C. burnetii) is the etiological agent of a Q fever-the re-emerging disease with considerable economic impact. Due to many similar symptoms with commonly occurring infections, its clinical diagnosis is very difficult. Thus, a strong effort should be taken to raise the awareness and develop a robust strategy for an accurate diagnosis. The identification of specific C. burnetii biomarkers could be valuable for a sensitive and selective diagnosis of the disease. Herein, we described a workflow to identify immunoreactive proteins of C. burnetii with a high confidence. It is based on immunocapturing of bacterial antigens by biofunctionalized magnetic microspheres, followed by tandem mass spectrometry (MS/MS) identification. We detected dozens of previously reported antigens and proposed 15 novel biomarkers, which specificity was confirmed by in silico epitope analysis. Among them, the cardiolipin synthetase participating in the synthesis of cardiolipin was recognized. This biomarker could play a critical role in the early management of acute Q fever and prevention of Q fever endocarditis.

A novel biosensor based on serum antibody immobilization for rapid detection of viral antigens.

In this paper, we represent a label-free biosensor based on immobilization of serum antibodies for rapid detection of viral antigens. Human serum containing specific antibodies against Japanese encephalitis virus (JEV) was immobilized on a silanized surface of an interdigitated sensor via protein A/glutaraldehyde for electrical detection of JEV antigens. The effective immobilization of serum antibodies on the sensor surface was verified by Fourier transform infrared spectrometry and fluorescence microscopy. The signal of the biosensor obtained by the differential voltage converted from the change into non-Faradic impedance resulting from the specific binding of JEV antigens on the surface of the sensor. The detection analyzed indicates that the detection range of this biosensor is 1-10 µg/ml JEV antigens, with a detection limit of 0.75 µg/ml and that stable signals are measured in about 20 min. This study presents a useful biosensor with a high selectivity for rapid and simple detection of JEV antigens, and it also proposes the biosensor as a future diagnostic tool for rapid and direct detection of viral antigens in clinical samples for preliminary pathogenic screenings in the case of possible outbreaks.

Incidence of sperm-immobilizing antibodies in infertile women with past Chlamydia trachomatis infection.

PROBLEM: Among the risk factors for antisperm antibody production, inflammatory diseases of the genital tract are believed to play an important role. Chlamydia trachomatis infection is one of the most common sexually transmitted diseases. There are some reports suggesting that human sperm have antigens that cross-react immunologically with certain microbial antigens, such as C. trachomatis. However, this is still controversial. We performed a retrospective study to investigate the correlation between anti-chlamydial antibodies and sperm-immobilizing antibodies in infertile women. METHOD OF STUDY: Between January 2007 and March 2009, the presence of sperm-immobilizing antibodies was examined by the sperm immobilization test using sera from 273 infertile women. Anti-chlamydial antibodies (IgG and IgA) were examined to prove past C. trachomatis infection by ELISA using the same sera from infertile women. RESULTS: The overall incidence of sperm-immobilizing antibodies was 2.9% (8/273) in infertile women. The incidences of sperm-immobilizing antibodies were 6.4% (5/78) in cases with past C. trachomatis infection and 1.5% (3/195) in cases without past C. trachomatis infection. There was a significant difference between the two groups (P = 0.031). CONCLUSION: A significantly higher incidence of sperm-immobilizing antibodies was noted in infertile women with past C. trachomatis infection compared with that of those without past C. trachomatis infection. This is the first demonstration that C. trachomatis infection could play a role in the production of sperm-immobilizing antibodies in infertile women.

Visual sandwich immunoassay system on the basis of plasmon resonance scattering signals of silver nanoparticles.

In this contribution, we established a sandwich immunoassay system with a common spectrofluorometer to collect the plasmon resonance scattering (PRS) signals from silver nanoparticles (AgNPs) immunotargeted on glass slides. By taking the immunoreactions of goat antihuman IgG (Fc fragment specific) antibody (GAH-IgG), human immunoglobulin (H-IgG), and rabbit antihuman IgG (Fab fragment specific) antibody (RAH-IgG) as an example, we found that if a primary antibody (GAH-IgG) was first immobilized on the surface of glass slides and applied to capture target antigen (H-IgG), AgNPs-labeled secondary antibody (RAH-IgG) could be employed to detect the target antigen (H-IgG) by forming a sandwich immune complex on the surface of the glass slide. It was found that the PRS signals resulting from the AgNPs immunotargeted on the glass slides could be applied to the quantitative detection of H-IgG target antigen in the range of 10-1000 ng/mL with the limit of determination of 1.46 ng/mL (3sigma) under optimal conditions, which is sensitive and comparable with reported chemiluminescence immunoassays. With a dark-field microscope coupled with a spectral system, we measured the PRS features of single AgNPs immunotargeted on the glass slides, showing that the PRS of single nanoparticles might have potential applications in analytical chemistry. Further findings showed that the strong PRS signals from the AgNPs immunotargeted on the glass slides can be clearly seen and distinguished by naked eyes under the excitation of a common white light-emitting diode (LED) torch. Therefore, a visual PRS immunoassay system can be established easily with common glass slides and an LED torch.