Abstract
The Chlamydia pneumoniae genome‐encoded open reading frames Cpn0146, Cpn0147, and Cpn0308 were expressed as recombinant proteins for detecting C. pneumoniae‐specific antibodies in samples from three groups of individuals including 183 with C. pneumoniae‐associated respiratory infection (group I), 60 healthy blood donors (group II), and 32 with no known respiratory infection (group III). The recombinant Cpn0146 was recognized by 71 (38.8% positive recognition rate), 15 (25%) and 1 (3.1%), Cpn0147 by 75 (40.9%), 14 (23.3%), and 2 (6.3%), and Cpn0308 by 82 (44.8%), 16 (26.7%), and 0 (0%) samples from groups I, II, and III, respectively. The positive recognition rates with any of the three antigens were significantly higher in group I than those in groups II and III, suggesting that more individuals from group I were likely infected with C. pneumoniae. This conclusion was confirmed with a commercially available whole organism‐based ELISA kit (Savyon Diagnostics Ltd., Ashdod, Israel), which detected C. pneumoniae antibodies in 98 (64.1%), 26 (43.3%), and 4 (12.5%) samples from group I, II, and III, respectively. Comparing to the commercial kit, the recombinant antigen‐based detection assays displayed >97% of detection specificity and >87% of sensitivity, suggesting that these recombinant antigens can be considered alternative tools for aiding in serodiagnosis of C. pneumoniae infection. J. Clin. Lab. Anal. 24:55–61, 2010. © 2010 Wiley‐Liss, Inc.
Keywords: Chlamydophila pneumoniae, serodiagnosis, recombinant antigens, ELISA
REFERENCES
- 1. Grayston JT, Kuo CC, Wang SP, Altman J. A new Chlamydia psittaci strain, TWAR, isolated in acute respiratory tract infections. N Engl J Med 1986;315:161–168. [DOI] [PubMed] [Google Scholar]
- 2. Kuo C‐C, Jackson LA, Campbell LA, et al. Chlamydia pneumoniae (TWAR). Clin Microbiol Rev 1995;8:451–461. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. Kumar S, Hammerschlag MR. Acute respiratory infection due to Chlamydia pneumoniae: Current status of diagnostic methods. Clin Infect Dis 2007;44:568–576. [DOI] [PubMed] [Google Scholar]
- 4. Sessa R, Nicoletti M, Di Pietro M, et al. Chlamydia pneumoniae and atherosclerosis: Current state and future prospectives. Int J Immunopathol Pharmacol 2009;22:9–14. [DOI] [PubMed] [Google Scholar]
- 5. Boman J, Hammerschlag MR. Chlamydia pneumoniae and atherosclerosis: Critical assessment of diagnostic methods and relevance to treatment studies. Clin Micro 2002;15:1–20. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6. Belland RJ, Ouellette SP, Gieffers J, Byrne GI. Chlamydia pneumoniae and atherosclerosis. Cell Microbiol 2004;6:117–127. [DOI] [PubMed] [Google Scholar]
- 7. Verkooyen RP, Willemse D, Hiep‐van Casteren SC, et al. Evaluation of PCR, culture, and serology for diagnosis of Chlamydia pneumoniae respiratory infections. J Clin Microbiol 1998;36:2301–2307. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. Paldanius M, Bloigu A, Leinonen M, Saikku P. Measurement of Chlamydia pneumoniae‐specific immunoglobulin A (IgA) antibodies by the microimmunofluorescence (MIF) method: Comparison of seven fluorescein‐labeled anti‐human IgA conjugates in an in‐house MIF test using one commercial MIF and one enzyme immunoassay kit. Clin Diagn Lab Immunol 2003;10:8–12. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. Wong YK, Sueur JM, Fall CH, Orfila J, Ward ME. The species specificity of the microimmunofluorescence antibody test and comparisons with a time resolved fluoroscopic immunoassay for measuring IgG antibodies against Chlamydia pneumoniae . J Clin Pathol 1999;52:99–102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10. Bennedsen M, Berthelsen L, Lind I. Performance of three microimmunofluorescence assays for detection of Chlamydia pneumoniae immunoglobulin M, G, and A antibodies. Clin Diagn Lab Immunol 2002;9:833–839. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. Miyashita N, Ouchi K, Kawasaki K, et al. Evaluation of enzyme‐linked immunosorbent assay for Chlamydophila pneumoniae‐specific immunoglobulin M in acute respiratory tract infection. Respirology 2008;13:299–302. [DOI] [PubMed] [Google Scholar]
- 12. Kutlin A, Tsumura N, Emre U, Roblin PM, Hammerschlag MR. Evaluation of Chlamydia immunoglobulin M (IgM), IgG, and IgA rELISAs Medac for diagnosis of Chlamydia pneumoniae infection. Clin Diagn Lab Immunol 1997;4:213–216. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13. Luo J, Liu G, Zhong Y, Jia T, Zhong G. Characterization of hypothetical proteins Cpn0146, 0147, 0284 & 0285 that are predicted to be in the Chlamydia pneumoniae inclusion membrane. BMC Microbiol 2007;15:7–38. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14. Luo J, Jia T, Flores R, Chen D, Zhong G. Hypothetical protein Cpn0308 is localized in the Chlamydia pneumoniae inclusion membrane. Infect Immun 2007;75:497–503. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. Harry T, Theophil S, Julian G. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci 1979;76:4350–4354. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. Masse J. Day‐to‐day variation in iron‐status indices. Am J Clinical Nutrition 1992;55:1187–1189. [DOI] [PubMed] [Google Scholar]
- 17. Kalay N, Kutukoglu I, Ozdogru I, et al. The relationship between Chlamydophila pneumoniae IgG titer and coronary atherosclerosis. Cardiol J 2008;15:245–251. [PubMed] [Google Scholar]
- 18. Numazaki K, Ikebe T, Chiba S. Detection of serum antibodies against Chlamydia pneumoniae by ELISA. FEMS Immunol Med Microbiol 1996;14:179–183. [DOI] [PubMed] [Google Scholar]
- 19. Sueur JM, Beaumont K, Cabioch T, Orfila J, Betsou F. Diagnostic value of an ELISA using a recombinant 54 kDa species‐specific protein from Chlamydia pneumoniae . Clin Microbiol Infect 2006;12:470–477. [DOI] [PubMed] [Google Scholar]
- 20. Kazemi B, Seyed N, Bandehpour M, Sharifnia Z, Pakzad P. Cloning, expression and purification of truncated Chlamydia trachomatis outer membrane protein 2 (omp2) and its application in an ELISA assay. Iran J Immunol 2008;5:148–155. [PubMed] [Google Scholar]
- 21. Wolf K, Fischer E, Mead D, et al. Chlamydia pneumoniae major outer membrane protein is a surface‐exposed antigen that elicits antibodies primarily directed against conformation‐dependent determinants. Infect Immun 2001;69:3082–3091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22. Montigiani S, Falugi F, Scarselli M, et al. Genomic approach for analysis of surface proteins in Chlamydia pneumoniae . Infect Immun 2002;70:368–379. [DOI] [PMC free article] [PubMed] [Google Scholar]