PMID- 8877384
OWN - NLM
STAT- MEDLINE
DCOM- 19970221
LR  - 20191101
IS  - 0948-6143 (Print)
IS  - 0948-6143 (Linking)
VI  - 106
IP  - 2
DP  - 1996 Aug
TI  - Fluorescence in situ hybridization with cosmid clones for the detection of human 
      cytomegalovirus DNA in peripheral blood leukocytes.
PG  - 229-34
AB  - Radioactive in situ hybridization techniques or enzymatic detection procedures of 
      hapten-modified human cytomegalovirus (HCMV) probes have been widely used for 
      studying the infection of peripheral blood leukocytes with HCMV. This report 
      describes significant improvements in terms of signal resolution which can be 
      obtained by applying a highly sensitive fluorescence in situ hybridization (FISH) 
      technique in conjunction with a large subgenomic HCMV DNA probe. Three cosmid 
      clones spanning 119.1 kb of the HCMV genome (230 kb) were used to construct the 
      digoxigenin-11-dUTP-labeled probe which was found to be superior to a total HCMV 
      probe representing the entire genome. Crucial hybridization parameters were 
      analyzed systematically in order to ensure optimal resolution power and 
      sensitivity. The protocol was successfully applied to HCMV-infected fibroblasts 
      and peripheral blood leukocytes of 12 transplant patients and unambiguously 
      facilitated the precise intracellular localization of HCMV genomes in infected 
      cells. Because of its excellent resolution properties, accompanied by the virtual 
      absence by any background staining, we recommend the use of this protocol as a 
      sensitive approach for further virological analyses of the interactions between 
      HCMV and peripheral blood leukocytes at the single-cell level.
FAU - Hackstein, H
AU  - Hackstein H
AD  - Institute of Immunology and Transfusion Medicine, University of Lubeck School of 
      Medicine, Germany.
FAU - Jahn, G
AU  - Jahn G
FAU - Kirchner, H
AU  - Kirchner H
FAU - Bein, G
AU  - Bein G
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Histochem Cell Biol
JT  - Histochemistry and cell biology
JID - 9506663
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Satellite)
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Cloning, Molecular
MH  - Cosmids
MH  - Cytomegalovirus/*isolation & purification
MH  - Cytomegalovirus Infections/virology
MH  - DNA Probes
MH  - DNA, Satellite
MH  - DNA, Viral/*analysis
MH  - Fibroblasts/virology
MH  - Genetic Techniques
MH  - Humans
MH  - Immunoenzyme Techniques
MH  - *In Situ Hybridization, Fluorescence
MH  - Leukocytes/*virology
MH  - Organ Transplantation
MH  - Sensitivity and Specificity
EDAT- 1996/08/01 00:00
MHDA- 1996/08/01 00:01
CRDT- 1996/08/01 00:00
PHST- 1996/08/01 00:00 [pubmed]
PHST- 1996/08/01 00:01 [medline]
PHST- 1996/08/01 00:00 [entrez]
AID - 10.1007/BF02484405 [doi]
PST - ppublish
SO  - Histochem Cell Biol. 1996 Aug;106(2):229-34. doi: 10.1007/BF02484405.