Development of a Human Herpesvirus 6 Species-Specific Immunoblotting Assay
| Autor: | Masaru Ihira, Ken Sugata, Tetsushi Yoshikawa, Akane Ohta, Yukihiro Nishiyama, Yuki Higashimoto, Dharam V. Ablashi, Daniel L. Peterson, Paolo Lusso, Yoshizo Asano |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Microbiology (medical) Adolescent medicine.drug_class Herpesvirus 6 Human viruses Blotting Western Antibodies Viral Monoclonal antibody Peripheral blood mononuclear cell Virus law.invention Young Adult Antigen law Virology Immunoblot Analysis Exanthema Subitum medicine Humans Child Antigens Viral Cells Cultured Aged biology Infant virus diseases Middle Aged biology.organism_classification Molecular biology Child Preschool DNA Viral Leukocytes Mononuclear Recombinant DNA biology.protein Female Human herpesvirus 6 Antibody |
| Zdroj: | Journal of Clinical Microbiology. 50:1245-1251 |
| ISSN: | 1098-660X 0095-1137 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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