protein microarray immunoassay for the serological evaluation of the antibody response in vertically transmitted infections

• Published in: European journal of clinical microbiology & infectious diseases 2009-09, Vol.28 (9), p.1067-1075
• Main Authors: Ardizzoni, A, Capuccini, B, Baschieri, M. C, Orsi, C. F, Rumpianesi, F, Peppoloni, S, Cermelli, C, Meacci, M, Crisanti, A, Steensgaard, P, Blasi, E
• Format: Article
• Language: English
• Subjects: Antibodies, Bacterial - blood; Antibodies, Protozoan - blood; Antibodies, Viral - blood; Antibody response; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Chlamydia trachomatis; Communicable Diseases - diagnosis; Communicable Diseases - immunology; Cytomegalovirus; Enzyme-linked immunosorbent assay; Female; Herpes simplex virus; Humans; Immobilization; Immunofluorescence; Immunoglobulin G; Immunoglobulin G - blood; Immunoglobulin M; Immunoglobulin M - blood; Infection; Infectious Disease Transmission, Vertical; Infectious diseases; Internal Medicine; Medical Microbiology; Medical sciences; Microscopes; Pathogens; Pregnancy; Protein Array Analysis - methods; Rubella; Rubella virus; Sensitivity and Specificity; Sexually transmitted diseases; STD; Toxoplasma gondii; Varicella; Varicella-zoster virus
• ISSN: 0934-9723; 1435-4373
• DOI: 10.1007/s10096-009-0748-y

The detection of specific serum antibodies is mainly achieved by enzyme-linked immunosorbent assay (ELISA). Here, we describe the setting up of a microarray-based serological assay to screen for IgG and IgM against vertically transmitted pathogens (Toxoplasma gondii, rubella virus, cytomegalovirus, herpes simplex virus types 1 and 2, varicella zoster virus, Chlamydia trachomatis). The test, accommodated onto a restricted area of a microscope slide, consists of: (a) the immobilization of antigens and human IgG and IgM antibody dilution curves, laid down in an orderly manner; (b) addition of serum samples; (c) detection of antigen-serum antibodies complexes by indirect immunofluorescence. The IgG and IgM curves provide an internal calibration system for the interpolation of the signals from the single antigens. The test was optimized in terms of spotting conditions and processing protocol. The detection limit was 400 fg for the IgG assay and 40 fg for the IgM assay; the analytical specificity was >98%. The clinical sensitivity returned an average value of 78%, the clinical specificity was >96%, the predictive values were >73%, and the efficiency was >88%. The results obtained make this test a promising tool, suitable for introduction in the clinical diagnostic routine of vertically transmitted infections, in parallel (and in future as an alternative) to ELISA.