Development of recombinant antigen array for simultaneous detection of viral antibodies.
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.
Pediatric Research Institute, Qilu Children's Hospital of Shandong University, Ji'nan, China.
Yi
Y
Liu
Fengling
F
Yu
Haiyan
H
Huang
Jinxiang
J
Han
D000293:Adolescent; D000328:Adult; D000368:Aged; D000914:Antibodies, Viral / Q000097:blood / Q000134:cerebrospinal fluid; D000915:Antibody Affinity; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000097:blood / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D003586:Cytomegalovirus Infections / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D005260:Female; D006558:Herpes Genitalis / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D006561:Herpes Simplex / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000134:cerebrospinal fluid; D007075:Immunoglobulin M / Q000097:blood / Q000134:cerebrospinal fluid; D007223:Infant; D008297:Male; D008875:Middle Aged; D010750:Phosphoproteins / Q000097:blood / Q000276:immunology; D040081:Protein Array Analysis / Q000379:methods*; D012409:Rubella / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D014760:Viral Fusion Proteins / Q000097:blood / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D010750:Phosphoproteins; D014760:Viral Fusion Proteins