title: Development of a human herpesvirus 6 species-specific immunoblotting assay.
abstract: In order to assess the full spectrum of human herpesvirus 6A (HHV-6A)- and HHV-6B-associated diseases, we sought to develop an HHV-6 species-specific serological assay based on immunoblot analysis. The immunodominant proteins encoded by open reading frame U11, p100 for HHV-6A (strain U1102) and 101K for HHV-6B (strain Z29), were selected to generate virus species-specific antigens. Recombinant p100 and 101K were produced in a prokaryotic expression system. The expression of these proteins was confirmed by using anti-His tag and 101K-specific monoclonal antibodies. HHV-6 species-specific antibodies were detected by immunoblotting in patient sera. Eighty-seven serum samples obtained from various subjects were utilized to determine the reliability of the method for clinical use. Ten of twelve exanthem subitum convalescent-phase sera reacted exclusively with 101K, whereas none of twelve acute-phase sera reacted with either protein. Two of three sera collected from HHV-6A-infected patients reacted with p100 and 101K. Although all five acute and convalescent-phase sera obtained from transplant recipients reacted exclusively with 101K, two of six convalescent-phase sera obtained from patients with drug-induced hypersensitivity syndrome reacted with both p100 and 101K. Of 38 sera obtained from healthy adults, 31 were positive for 101K antibody, while 4 reacted with both proteins. However, PCR analysis of peripheral blood mononuclear cells and saliva from these subjects did not detect HHV-6A DNA. In conclusion, this novel serological assay based on immunoblot analysis using recombinant HHV-6A p100 and HHV-6B 101K allowed us to discriminate between HHV-6A- and HHV-6B-specific antibodies.
authors.affiliation: Department of Pediatrics, Fujita Health University School of Medicine, Toyoake, Aichi, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Higashimoto
authors.forename: Akane
authors.initials: A
authors.lastname: Ohta
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Nishiyama
authors.forename: Masaru
authors.initials: M
authors.lastname: Ihira
authors.forename: Ken
authors.initials: K
authors.lastname: Sugata
authors.forename: Yoshizo
authors.initials: Y
authors.lastname: Asano
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Peterson
authors.forename: Dharam V
authors.initials: DV
authors.lastname: Ablashi
authors.forename: Paolo
authors.initials: P
authors.lastname: Lusso
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Yoshikawa
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology*; D015153:Blotting, Western; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000097:blood; D005077:Exanthema Subitum / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D005260:Female; D015654:Herpesvirus 6, Human / Q000276:immunology*; D006801:Humans; D007223:Infant; D007963:Leukocytes, Mononuclear / Q000821:virology; D008297:Male; D008875:Middle Aged; D055815:Young Adult
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral