PMID- 8349756
OWN - NLM
STAT- MEDLINE
DCOM- 19930914
LR  - 20210526
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 31
IP  - 7
DP  - 1993 Jul
TI  - Effects of fixation and varying target length on the sensitivity of polymerase 
      chain reaction for detection of human T-cell leukemia virus type I proviral DNA 
      in formalin-fixed tissue sections.
PG  - 1799-803
AB  - In this study, the fixation condition most suitable for maintaining the 
      sensitivity of the polymerase chain reaction (PCR) was investigated by using the 
      alpha-tubulin gene sequence, and the PCR procedure most effective for detecting 
      human T-cell leukemia virus type I (HTLV-I) proviral sequences in fixed, embedded 
      tissues of adult T-cell leukemia patients was explored. First, the sensitivity of 
      the PCR targeting a 286-bp alpha-tubulin sequence was studied in tissue sections 
      fixed in several fixatives for various periods at 25 or 4 degrees C. For 
      histological examination, fixation with 10% buffered formalin at a lower 
      temperature for a shorter period was found to be preferable to retain the 
      sensitivity. And the HTLV-I sequence was detected in only 7 of 18 specimens 
      (38.9%) when the 374-bp sequence of the gag region was targeted, but the rate 
      increased to 77.8% (14 of 18 specimens) when the length of the target sequence 
      was reduced to 120 bp within the same region. Therefore, the one-round PCR 
      targeting a shorter sequence is preferable for application of PCR to archival 
      fixed tissue specimens, the fixation condition of which may not be ideal for DNA 
      preservation.
FAU - Honma, M
AU  - Honma M
AD  - Department of Neurological Sciences, Tohoku University School of Medicine, 
      Sendai, Japan.
FAU - Ohara, Y
AU  - Ohara Y
FAU - Murayama, H
AU  - Murayama H
FAU - Sako, K
AU  - Sako K
FAU - Iwasaki, Y
AU  - Iwasaki Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (DNA, Viral)
RN  - 0 (Fixatives)
RN  - 0 (Tubulin)
RN  - 1HG84L3525 (Formaldehyde)
SB  - IM
GS  - gag
MH  - Base Sequence
MH  - DNA, Viral/*genetics/isolation & purification
MH  - Evaluation Studies as Topic
MH  - Fixatives
MH  - Formaldehyde
MH  - Genes, gag
MH  - Human T-lymphotropic virus 1/*genetics/isolation & purification
MH  - Humans
MH  - Leukemia-Lymphoma, Adult T-Cell/microbiology
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction/*methods/statistics & numerical data
MH  - Proviruses/genetics/isolation & purification
MH  - Sensitivity and Specificity
MH  - Temperature
MH  - Tubulin/genetics
PMC - PMC265635
EDAT- 1993/07/01 00:00
MHDA- 1993/07/01 00:01
PMCR- 1993/07/01
CRDT- 1993/07/01 00:00
PHST- 1993/07/01 00:00 [pubmed]
PHST- 1993/07/01 00:01 [medline]
PHST- 1993/07/01 00:00 [entrez]
PHST- 1993/07/01 00:00 [pmc-release]
AID - 10.1128/jcm.31.7.1799-1803.1993 [doi]
PST - ppublish
SO  - J Clin Microbiol. 1993 Jul;31(7):1799-803. doi: 10.1128/jcm.31.7.1799-1803.1993.