PMID- 16254342
OWN - NLM
STAT- MEDLINE
DCOM- 20060127
LR  - 20181113
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 79
IP  - 22
DP  - 2005 Nov
TI  - Laser-capture microdissection: refining estimates of the quantity and 
      distribution of latent herpes simplex virus 1 and varicella-zoster virus DNA in 
      human trigeminal Ganglia at the single-cell level.
PG  - 14079-87
AB  - There remains uncertainty and some controversy about the percentages and types of 
      cells in human sensory nerve ganglia that harbor latent herpes simplex virus 1 
      (HSV-1) and varicella-zoster virus (VZV) DNA. We developed and validated 
      laser-capture microdissection and real-time PCR (LCM/PCR) assays for the presence 
      and copy numbers of HSV-1 gG and VZV gene 62 sequences in single cells recovered 
      from sections of human trigeminal ganglia (TG) obtained at autopsy. Among 970 
      individual sensory neurons from five subjects, 2.0 to 10.5% were positive for 
      HSV-1 DNA, with a median of 11.3 copies/positive cell, compared with 0.2 to 1.5% 
      of neurons found to be positive by in situ hybridization (ISH) for HSV-1 
      latency-associated transcripts (LAT), the classical surrogate marker for HSV 
      latency. This indicates a more pervasive latent HSV-1 infection of human TG 
      neurons than originally thought. Combined ISH/LCM/PCR assays revealed that the 
      majority of the latently infected neurons do not accumulate LAT to detectable 
      levels. We detected VZV DNA in 1.0 to 6.9% of individual neurons from 10 
      subjects. Of the total 1,722 neurons tested, 4.1% were VZV DNA positive, with a 
      median of 6.9 viral genomes/positive cell. After removal by LCM of all visible 
      neurons on a slide, all surrounding nonneuronal cells were harvested and assayed: 
      21 copies of HSV-1 DNA were detected in approximately 5,200 nonneuronal cells, 
      while nine VZV genomes were detected in approximately 14,200 nonneuronal cells. 
      These data indicate that both HSV-1 and VZV DNAs persist in human TG primarily, 
      if not exclusively, in a moderate percentage of neuronal cells.
FAU - Wang, Kening
AU  - Wang K
AD  - Medical Virology Section, Laboratory of Clinical Infectious Diseases, National 
      Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892, USA. 
      kwang@niaid.nih.gov
FAU - Lau, Tsz Y
AU  - Lau TY
FAU - Morales, Melissa
AU  - Morales M
FAU - Mont, Erik K
AU  - Mont EK
FAU - Straus, Stephen E
AU  - Straus SE
LA  - eng
GR  - Intramural NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Intramural
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Viral)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Base Sequence
MH  - DNA Primers
MH  - DNA, Viral/genetics/*isolation & purification/ultrastructure
MH  - Gene Expression Regulation, Viral
MH  - Herpesvirus 1, Human/*genetics/isolation & purification
MH  - Herpesvirus 3, Human/*genetics/isolation & purification
MH  - Humans
MH  - Lasers
MH  - Microdissection/methods
MH  - Polymerase Chain Reaction
MH  - RNA, Viral/genetics/isolation & purification
MH  - Trigeminal Ganglion/*virology
PMC - PMC1280223
EDAT- 2005/10/29 09:00
MHDA- 2006/01/28 09:00
PMCR- 2005/11/01
CRDT- 2005/10/29 09:00
PHST- 2005/10/29 09:00 [pubmed]
PHST- 2006/01/28 09:00 [medline]
PHST- 2005/10/29 09:00 [entrez]
PHST- 2005/11/01 00:00 [pmc-release]
AID - 79/22/14079 [pii]
AID - 1192-05 [pii]
AID - 10.1128/JVI.79.22.14079-14087.2005 [doi]
PST - ppublish
SO  - J Virol. 2005 Nov;79(22):14079-87. doi: 10.1128/JVI.79.22.14079-14087.2005.