Development of a human herpesvirus 6 species-specific immunoblotting assay

Yuki Higashimoto, Akane Ohta, Yukihiro Nishiyama, Masaru Ihira, Ken Sugata, Yoshizo Asano, Daniel L. Peterson, Dharam V. Ablashi, Paolo Lusso, Tetsushi Yoshikawa

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1245-1251
Number of pages7
JournalJournal of clinical microbiology
Volume50
Issue number4
DOIs
Publication statusPublished - 01-04-2012

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Human Herpesvirus 6
Immunoblotting
Serum
Antibodies
Proteins
Drug Hypersensitivity Syndrome
Exanthema
Saliva
Open Reading Frames
Blood Cells

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

Cite this

Higashimoto, Yuki ; Ohta, Akane ; Nishiyama, Yukihiro ; Ihira, Masaru ; Sugata, Ken ; Asano, Yoshizo ; Peterson, Daniel L. ; Ablashi, Dharam V. ; Lusso, Paolo ; Yoshikawa, Tetsushi. / Development of a human herpesvirus 6 species-specific immunoblotting assay. In: Journal of clinical microbiology. 2012 ; Vol. 50, No. 4. pp. 1245-1251.
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Higashimoto, Y, Ohta, A, Nishiyama, Y, Ihira, M, Sugata, K, Asano, Y, Peterson, DL, Ablashi, DV, Lusso, P & Yoshikawa, T 2012, 'Development of a human herpesvirus 6 species-specific immunoblotting assay', Journal of clinical microbiology, vol. 50, no. 4, pp. 1245-1251. https://doi.org/10.1128/JCM.05834-11

Development of a human herpesvirus 6 species-specific immunoblotting assay. / Higashimoto, Yuki; Ohta, Akane; Nishiyama, Yukihiro; Ihira, Masaru; Sugata, Ken; Asano, Yoshizo; Peterson, Daniel L.; Ablashi, Dharam V.; Lusso, Paolo; Yoshikawa, Tetsushi.

In: Journal of clinical microbiology, Vol. 50, No. 4, 01.04.2012, p. 1245-1251.

Research output: Contribution to journalArticle

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T1 - Development of a human herpesvirus 6 species-specific immunoblotting assay

AU - Higashimoto, Yuki

AU - Ohta, Akane

AU - Nishiyama, Yukihiro

AU - Ihira, Masaru

AU - Sugata, Ken

AU - Asano, Yoshizo

AU - Peterson, Daniel L.

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AU - Lusso, Paolo

AU - Yoshikawa, Tetsushi

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