Abstract
Protein microarrays have been developed to study antibody reactivity against a large number of antigens, demonstrating extensive perspective for clinical application. We developed a viral antigen array by spotting four recombinant antigens and synthetic peptide, including glycoprotein G of herpes simplex virus (HSV) type 1 and 2, phosphoprotein 150 of cytomegalovirus (CMV), Rubella virus (RV) core plus glycoprotein E1 and E2 as well as a E1 peptide with the optimal concentrations on activated glass slides to simultaneously detect IgG and IgM against HSV1, HSV2, CMV and RV in clinical specimens of sera and cerebrospinal fluids (CSFs). The positive reference sera were initially used to measure the sensitivity and specificity of the array with the optimal conditions. Then clinical specimens of 144 sera and 93 CSFs were tested for IgG and IgM antibodies directed against HSV1, HSV2, CMV and RV by the antigen array. Specificity of the antigen array for viral antibodies detection was satisfying compared to commercial ELISA kits but sensitivity of the array varied relying on quality and antigenic epitopes of the spotting antigens. In short, the recombinant antigen array has potential to simultaneous detect multiple viral antibodies using minute amount (3 µl) of samples, which holds the particularly advantage to detect viral antibodies in clinical CSFs being suspicious of neonatal meningitis and encephalitis.
Highlights
Protein microarray, a high-throughput and powerful detection tool in medical research, is characterized methodologically as that of gene microarray, including robotic printing of probe targets, hybridization with samples of interest, and detection of interaction by high-resolution scanning and image analysis [1,2]
We established an antigen microarray based on Enzyme-linked immunosorbent assays (ELISAs) assay and took ELISA as a gold standard for comparison of two methods in this study
The ELISA kits used were made from whole virus lysate
Summary
A high-throughput and powerful detection tool in medical research, is characterized methodologically as that of gene microarray, including robotic printing of probe targets, hybridization with samples of interest, and detection of interaction by high-resolution scanning and image analysis [1,2]. Protein microarrays have been applied to analyze serum and other biological fluids in different contexts including protein-protein interaction, cancer profiling, autoimmune and infectious diseases [3,4,5,6,7]. Determination of the viral infection is currently depended on virus isolation, antigen measurement, nucleic acid detection and serological methods [10,11,12]. Enzyme-linked immunosorbent assays (ELISAs) are exceptionally sensitive and specific for detecting the concentration of proteins in complex biological fluids of blood, cerebrospinal fluid and others. Protein microarrays have been developed to detect antibodies against infectious viruses by using crude extracts, semi-purified microbial antigens, and recombinant proteins with higher concordance rates between microarray assays and ELISAs [13,14,15]. The main advantage is simultaneous determination of multi-antibodies from 3 mL of serum or cerebrospinal fluid (CSF)
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