Cycling probe-based real-time PCR for the detection of Human herpesvirus 6A and B

Masaru Ihira, Ayumi Yamaki, Yuri Kato, Yuki Higashimoto, Yoshiki Kawamura, Tetsushi Yoshikawa

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Human herpesvirus 6 (HHV-6) is classified as two distinct species: HHV-6A and B. HHV-6B infection can cause several clinical manifestations in transplant recipients including encephalitis, bone marrow suppression, and pneumonitis. In contrast to HHV-6B, the clinical features of HHV-6A infection remain largely undefined. Herein, we developed a multiplex cycling probe real-time PCR that discriminated between HHV-6A and HHV-6B. The assay was HHV-6-specific and no cross amplification was demonstrated for other herpesviruses. Moreover, the assay had a broad, linear dynamic range of detection between 1 and 106 copies of viral DNA. The quantification of HHV-6A DNA was suppressed by an excess amount of HHV-6B DNA (1 × 106 copies/tube) in the multiplex PCR assay; however, 1 × 106 copies/tube of HHV-6A DNA did not affect the quantification of 1 × 104 copies/tube of HHV-6B DNA. To determine the reliability of the assay for analysis of clinical specimens, DNAs extracted from the peripheral blood of hematopoietic stem cell transplant recipients were assayed using our multiplex real-time PCR versus the standard TaqMan PCR. Strong correlations were demonstrated between the two different assay systems for both HHV-6A (R2 = 0.913) and HHV-6B (R2 = 0.909). Therefore, our multiplex HHV-6 species-specific cycling probe real-time PCR is useful for evaluating the precise copy numbers of HHV-6A and B in transplant recipients. However, as HHV-6A copy numbers was affected by presence of high copies of HHV-6B DNA (1 × 106 copies/tube), it may be difficult to measure precise copy numbers of HHV-6A in inherited chromosomally integrated HHV-6B patient. J. Med. Virol. 88:1628–1635, 2016.

Original languageEnglish
Pages (from-to)1628-1635
Number of pages8
JournalJournal of Medical Virology
Volume88
Issue number9
DOIs
Publication statusPublished - 01-09-2016

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All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases

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