Serodiagnosis of infectious diseases with antigen microarrays.
OBJECTIVE
To generate protein microarrays by printing microbial antigens on slides to enable the simultaneous determination in human sera of antibodies directed against Toxoplasma gondii, rubella virus, cytomegalovirus and herpes simplex virus (HSV) types 1 and 2.


RESULTS
Antigens were printed on activated glass slides using high-speed robotics. The slides were incubated with serum samples and subsequently with fluorescently labelled secondary antibodies. Human IgG and IgM bound to the printed antigens were detected using confocal scanning microscopy and quantified with internal calibration curves. The microarray assay could detect as little as 0.5 pg of both IgG and IgM bound onto the glass surface. Precision profiles ranged from 1.7 to 18.5% for all the antigens. Microarrays and commercial ELISAs were utilized to detect serum antibodies against the ToRCH antigens in a panel of characterized human sera. Overall >80% concordance was obtained between microarray and ELISA kits in the classification of sera.


CONCLUSIONS
These results indicate that the microarray is a suitable assay format for the serodiagnosis of infectious diseases.


CONCLUSIONS
Antigen microarrays can be optimized for clinical use, their performance is equivalent to ELISA but they offer significant advantages in throughput, convenience and cost.
Bacarese-Hamilton
T
T
Department of Biology, Imperial College of Science, Technology and Medicine, London, UK.
Mezzasoma
L
L
Ardizzoni
A
A
Bistoni
F
F
Crisanti
A
A
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D000953:Antigens, Protozoan / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D003587:Cytomegalovirus / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000175:diagnosis; D004797:Enzyme-Linked Immunosorbent Assay; D006561:Herpes Simplex / Q000175:diagnosis; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D040081:Protein Array Analysis / Q000379:methods*; D015203:Reproducibility of Results; D012698:Serologic Tests / Q000379:methods*; D018139:Simplexvirus / Q000276:immunology; D014122:Toxoplasma / Q000276:immunology; D014123:Toxoplasmosis / Q000175:diagnosis*; D014777:Virus Diseases / Q000175:diagnosis*
D000913:Antibodies, Protozoan; D000914:Antibodies, Viral; D000953:Antigens, Protozoan; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M