PMID- 22825817
OWN - NLM
STAT- MEDLINE
DCOM- 20121119
LR  - 20120724
IS  - 1096-9071 (Electronic)
IS  - 0146-6615 (Linking)
VI  - 84
IP  - 9
DP  - 2012 Sep
TI  - Development of quantitative RT-PCR assays for detection of three classes of 
      HHV-6B gene transcripts.
PG  - 1388-95
LID - 10.1002/jmv.23350 [doi]
AB  - The monitoring of active human herpesvirus 6 (HHV-6) B infection is important for 
      distinguishing between the reactivation and latent state of the virus. The aim of 
      this present study is to develop a quantitative reverse transcription polymerase 
      chain reaction (RT-PCR) assay for diagnosis of active viral infection. Primers 
      and probes for in house quantitative RT-PCR methods were designed to detect the 
      three kinetic classes of HHV-6B mRNAs (U90, U12, U100). Stored PBMCs samples 
      collected from 10 patients with exanthem subitum (primary HHV-6B infection) and 
      15 hematopoietic stem cell transplant recipients with HHV-6B reactivation were 
      used to evaluate reliability for testing clinical samples. Excellent linearity 
      was obtained with high correlation efficiency between the diluted RNA 
      (1-100 ng/reaction) and C(t) value of each gene transcript. The U90 and U12 gene 
      transcripts were detected in all of the peripheral blood mononuclear cells 
      (PBMCs) samples collected in acute period of primary HHV-6B infection. Only one 
      convalescent PBMCs sample was positive for the U90 gene transcript. Additionally, 
      the reliability of HHV-6B quantitative RT-PCRs for diagnosis of viral 
      reactivation in hematopoietic transplant recipients was evaluated. Relative to 
      virus culture, U90 quantitative RT-PCR demonstrated the highest assay 
      sensitivity, specificity, positive predictive value, and negative predictive 
      value. Thus, this method could be a rapid and lower cost alternative to virus 
      culture, which is difficult to perform generally, for identifying active HHV-6B 
      infection.
CI  - Copyright (c) 2012 Wiley Periodicals, Inc.
FAU - Ihira, Masaru
AU  - Ihira M
AD  - Faculty of Clinical Engineering, Fujita Health University School of Health 
      Sciences, Toyoake, Aichi, Japan.
FAU - Enomoto, Yoshihiko
AU  - Enomoto Y
FAU - Kawamura, Yoshiki
AU  - Kawamura Y
FAU - Nakai, Hidetaka
AU  - Nakai H
FAU - Sugata, Ken
AU  - Sugata K
FAU - Asano, Yoshizo
AU  - Asano Y
FAU - Tsuzuki, Motohiro
AU  - Tsuzuki M
FAU - Emi, Nobuhiko
AU  - Emi N
FAU - Goto, Tatsunori
AU  - Goto T
FAU - Miyamura, Koichi
AU  - Miyamura K
FAU - Matsumoto, Kimikazu
AU  - Matsumoto K
FAU - Kato, Koji
AU  - Kato K
FAU - Takahashi, Yoshiyuki
AU  - Takahashi Y
FAU - Kojima, Seiji
AU  - Kojima S
FAU - Yoshikawa, Tetsushi
AU  - Yoshikawa T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Med Virol
JT  - Journal of medical virology
JID - 7705876
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Adult
MH  - Child
MH  - Child, Preschool
MH  - Exanthema Subitum/*diagnosis/virology
MH  - Female
MH  - *Genes, Viral
MH  - Hematopoietic Stem Cell Transplantation/adverse effects
MH  - Herpesvirus 6, Human/*genetics/physiology
MH  - Humans
MH  - Leukocytes, Mononuclear/virology
MH  - Limit of Detection
MH  - Male
MH  - Middle Aged
MH  - Molecular Diagnostic Techniques
MH  - RNA, Messenger/genetics
MH  - RNA, Viral/*genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sensitivity and Specificity
MH  - Transcription, Genetic
MH  - Virus Activation
MH  - Young Adult
EDAT- 2012/07/25 06:00
MHDA- 2012/12/10 06:00
CRDT- 2012/07/25 06:00
PHST- 2012/07/25 06:00 [entrez]
PHST- 2012/07/25 06:00 [pubmed]
PHST- 2012/12/10 06:00 [medline]
AID - 10.1002/jmv.23350 [doi]
PST - ppublish
SO  - J Med Virol. 2012 Sep;84(9):1388-95. doi: 10.1002/jmv.23350.