title: New sandwich-type enzyme-linked immunosorbent assay for human MxA protein in a whole blood using monoclonal antibodies against GTP-binding domain for recognition of viral infection.
abstract: OBJECTIVE
  To develop a clinically significant and practical enzyme-linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection.
  
  
  METHODS
  A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP-binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin.
  
  
  RESULTS
  This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0-100.0%), and rapidity (<1.5 h). The present ELISA had a sensitivity of 100% and a specificity of 100% for viral infection when compared to samples from healthy control and 87.1% and 90.9% when compared to samples from the bacterial infection group.
  
  
  CONCLUSIONS
  We have developed a new ELISA for measuring MxA protein in human whole blood using mAbs specific for the GTP-binding domain of MxA. This ELISA has analytical performance enough for routine clinical assay and can be used in detecting the possibility of viral infection.
authors.affiliation: Fuji Research Laboratories, Kyowa Medex Co., Ltd., Shizuoka, Japan.
authors.forename: Mizuho
authors.initials: M
authors.lastname: Kawamura
authors.forename: Akira
authors.initials: A
authors.lastname: Kusano
authors.forename: Akiko
authors.initials: A
authors.lastname: Furuya
authors.forename: Nobuo
authors.initials: N
authors.lastname: Hanai
authors.forename: Hideki
authors.initials: H
authors.lastname: Tanigaki
authors.forename: Akihito
authors.initials: A
authors.lastname: Tomita
authors.forename: Akira
authors.initials: A
authors.lastname: Horiguchi
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Nagata
authors.forename: Toshiko
authors.initials: T
authors.lastname: Itazawa
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Adachi
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Okabe
authors.forename: Toshio
authors.initials: T
authors.lastname: Miyawaki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kohno
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D001424:Bacterial Infections / Q000097:blood / Q000175:diagnosis; D001665:Binding Sites; D015415:Biomarkers / Q000097:blood; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D019204:GTP-Binding Proteins / Q000097:blood* / Q000276:immunology; D006801:Humans; D007223:Infant; D008297:Male; D051379:Mice; D064547:Myxovirus Resistance Proteins; D055550:Protein Stability; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000097:blood / Q000175:diagnosis*
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D019204:GTP-Binding Proteins