Use of recombinant chimeric antigens for the serodiagnosis of Mycoplasma pneumoniae infection.
In this paper, we have evaluated the diagnostic utility of three antigenic regions of the Mycoplasma pneumoniae P1, P30, and MPN456 gene products in order to replace the soluble, whole-cell bacterial extract in serological assays. Antigenic regions, being previously identified as B-cell epitopes, were used individually or assembled in a recombinant chimeric antigen by genetic engineering. Paired serum samples from 47 patients with M. pneumoniae infection and from 39 subjects with a clinical picture of atypical pneumonia but without a defined diagnosis of M. pneumoniae infection were included. Immunoglobulin G (IgG) antibodies against epitopes carried by recombinant antigens were measured by performing recombinant enzyme-linked immunosorbent assays (Rec-ELISAs). Rec-ELISA results were compared to those obtained by a commercial assay using the whole-cell Mycoplasma antigen. Our study demonstrates that all IgG Rec-ELISAs using recombinant antigens have better sensitivity with respect to the commercial assay. Furthermore, we show that the use of chimeric antigens improve the performance of the assays. The use of recombinant antigens is effective in distinguishing M. pneumoniae-infected patients from uninfected individuals and shows that immunoassays based on recombinant antigens could provide the basis for standardized commercial tests for the serodiagnosis of M. pneumoniae diseases.
Montagnani
F
F
Clinic of Infectious Diseases, University of Siena, Viale Bracci 16, 53100, Siena, Italy. montagnani2@unisi.it
De Paolis
F
F
Beghetto
E
E
Gargano
N
N
D000293:Adolescent; D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood* / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D007223:Infant; D009177:Mycoplasma pneumoniae / Q000276:immunology*; D011019:Pneumonia, Mycoplasma / Q000175:diagnosis* / Q000276:immunology; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D012698:Serologic Tests
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D018985:Epitopes, B-Lymphocyte; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins