title: Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of Chlamydophila pneumoniae IgA antibodies.
abstract: BACKGROUND
  Serologic diagnosis of Chlamydophila pneumoniae (Cpn) infection routinely involves assays for the presence of IgG and IgM antibodies to Cpn. Although IgA antibodies to Cpn have been found to be of interest in the diagnosis of chronic infections, their significance in serological diagnosis remains unclear. The microimmunofluorescence (MIF) test is the current method for the measurement of Cpn antibodies. While commercial enzyme linked immunosorbent assays (ELISA) have been developed, they have not been fully validated. We therefore evaluated and optimized a commercial ELISA kit, the SeroCP IgA test, for the detection of Cpn IgA antibodies.
  
  
  METHODS
  Serum samples from 94 patients with anti-Cpn IgG titers > or = 256 (study group) and from 100 healthy blood donors (control group) were tested for the presence of IgA antibodies to Cpn, using our in-house MIF test and the SeroCP IgA test. Two graph receiver operating characteristic (TG-ROC) curves were created to optimize the cut off given by the manufacturer.
  
  
  RESULTS
  The MIF and SeroCP IgA tests detected Cpn IgA antibodies in 72% and 89%, respectively, of sera from the study group, and in 9% and 35%, respectively, of sera from the control group. Using the MIF test as the reference method and the cut-off value of the ELISA test specified by the manufacturer for seropositivity and negativity, the two tests correlated in 76% of the samples, with an agreement of K = 0.54. When we applied the optimized cut-off value using TG-ROC analysis, 1.65, we observed better concordance (86%) and agreement (0.72) between the MIF and SeroCP IgA tests.
  
  
  CONCLUSIONS
  Use of TG-ROC analysis may help standardize and optimize ELISAs, which are simpler, more objective and less time consuming than the MIF test. Standardization and optimization of commercial ELISA kits may result in better performance.
authors.affiliation: Department of Microbiology and research laboratory Microorganismes et Pathologie Humaine, Habib Bourguiba Hospital of Sfax, Tunisia. Olfafrikha2005@yahoo.fr
authors.forename: Olfa
authors.initials: O
authors.lastname: Frikha-Gargouri
authors.forename: Radhouane
authors.initials: R
authors.lastname: Gdoura
authors.forename: Abir
authors.initials: A
authors.lastname: Znazen
authors.forename: Nozha
authors.initials: N
authors.lastname: Ben Arab
authors.forename: Jalel
authors.initials: J
authors.lastname: Gargouri
authors.forename: Mounir
authors.initials: M
authors.lastname: Ben Jemaa
authors.forename: Adnene
authors.initials: A
authors.lastname: Hammami
mesh_terms: D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood*; D016022:Case-Control Studies; D023521:Chlamydophila Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000276:immunology; D016993:Chlamydophila pneumoniae / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood*; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D016036:Seroepidemiologic Studies; D014416:Tunisia / Q000453:epidemiology
chemical_list: D000907:Antibodies, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G