Development of Protein Chip for Diagnosis of Chlamydia Pneumoniae

Background; The diagnosis of chlamydial infection is based on serology. The current gold standard of diagnosis is MIF(microimmunofluorescence), but this modality is subjective and time-consuming. Protein microarray with using a SPR(surface plasmon resonance) sensor has recently been suggested as a method for detecting infection. For developing a protein chip to diagnose chlamydial infection, EBs(elementary bodies) were immobilized on a gold chip and the interaction between an antibody for Chlamydophila pneumoniae and the EBs(elementary bodies) immobilized on the surface of the gold chip was measured by using an SPR sensor. Methods; For the surface antigen, the EBs of Chlamydophila pneumoniae LKK1 were purified. Charged arrays were prepared by using PDDA(polydiallyldimethylammonium chloride) which has a positive charge. After immobilization of the chlamydial EBs on the PDDA surface, the investigation of the surface was done with using atomic force microscopy. After the antibody for C. pneumoniae was applied on chip, we monitored the SPR wavelength-shift to detect any antigen-antibody interaction with using a self-assembled SPR sensor.Results; The chlamydial EBs on the positively charged PDDA were visible on the surface with using atomic force microscopy. The SPR wavelength increased after interaction of antibody for C. pneumoniae with the EBs immobilized on charged gold surface. The wavelength-shift was correlated with the concentration of antigens.Conclusion; The surface immobilization of EBs on the gold surface with the charged arrays was identified and the antigen-antibody interaction on the gold chip was detected via the SPR sensor. Further investigations are needed to apply this technique to the clinical field. (Tuberc Respir Dis 2006; 60: 412-418)

keywords: Chlamydophila pneumoniae, Protein array analysis, Chlamydophila pneumoniae, Protein array analysis

Reference

(1998) The role of atypical pathogens Chlamydia pneumoniae and Legionella pneumophila in respiratory infection,

(2002) Atypical pathogens in adult patients admitted with community-acquired pneumonia in Korea,

(1999) Chlamydia infections and heart disease linked through antigenic mimicry,

(2002) Comparison of individuals with and without specific IgA antibodies to Chlamydia pneumoniae,

(2001) Standardizing Chlamydia pneumoniae assays:recommendations from the centers for disease control and prevention and the laboratory centre for disease control,

(2003) Community-acquired pneumonia,

(2004) Molecular signature for diagnosis of infection:application of microarray technology,

(2002) Antigen microarrays for serodiagnosis of infectious diseases,

(2002) Surface plasmon resonance detection of serum antibodies against Salmonella enteritidis and Salmonella typhimurium,

10.

(2003) Immunochemical detection of Salmonella group B,D and E using an optical surface plasmon resonance biosensor,

11.

(2000) Printing proteins as microarrays for high-throughput function determination,

12.

(2003) Protein chip technology,

13.

(2003) Characterization of the first Korean isolate of a Chlamydia pneumoniae strain,

14.

(2003) Characterization of surface plasmon resonance wavelength by changes of protein concentration on protein chips,

15.

(2004) Comparison of quantitative and semiquantitative ELISA for IgG against C.pneumoniae to a MIF test for use with patients with respiratory tract infections,

16.

(2005) ELISA for antibodies against Chlamydia pneumoniae compared with MIF test with patients with chronic cough,

17.

(1999) Value of the polymerase chain reaction assay in noninvasive respiratory samples for diagnosis of community- acquired pneumonia,

18.

(2005) Proteomic applications of surface plasmon resonance biosensor:analysis of protein arrays,

19.

(2002) Microarray-based detection and genotyping of viral pathogens,

20.

(2005) Screening of specific antigens for SARS clinical diagnosis using a protein microarray,

21.

(2005) DNA microarray-based detection and identification of Chlamydia and Chlamydophila spp,

22.

(2002) Toward optimized antibody microarrays:a comparison of current microarray support materials,

23.

(1980) Surface properties of Chlamydia psittaci,

24.

(2003) Investigation of selection protein immobilization on charged protein array by wavelength interrogation-based SPR sensor,

25.

(2004) Analysis of protein interactions on protein arrays by a wavelength interrogation-based surface plasmon resonance biosensor,

26.

(2001) Surface plasmon resonance:towards an understanding of the mechanisms of biological molecular recognition,

27.

(2002) Elimination of ingredients effect to improve the detection of anti HIV-1 p24 antibody in human serum using SPR apparatus,

바로가기메뉴

Article Contents

Vol.60 No.4

Development of Protein Chip for Diagnosis of Chlamydia Pneumoniae

Abstract

Reference

Tuberculosis & Respiratory Diseases