Medical College of Wisconsin
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Detection of Epstein-Barr virus EBER sequence in post-transplant lymphoma patients with DNA dendrimers. J Virol Methods 1999 Feb;77(2):153-63

Date

03/26/1999

Pubmed ID

10092139

DOI

10.1016/s0166-0934(98)00149-9

Scopus ID

2-s2.0-0033042328 (requires institutional sign-in at Scopus site)   17 Citations

Abstract

The EBER RNAs are the most numerous viral transcripts in latently infected lymphocytes in healthy individuals and also in the tumor cells of Epstein Barr virus (EBV)-associated malignancies. A rise in EBV load in peripheral blood has been associated with the onset of post-transplant lymphoproliferative disease (PTLD) in immunocompromised patients. Treatment of PTLD with adoptive immunotherapy has made the rapid and accurate determination of EBV load essential. The relationship between EBV load and other EBV-associated malignancies, like Hodgkin's disease or AIDS-associated lymphoma, is unknown. In order to define viral load based on the number of EBV-infected cells in the peripheral blood, we developed a method which combines cellular dilution of peripheral blood mononuclear cells with the direct detection of EBER-1 RNA with DNA dendrimers. DNA dendrimers are large scaffolds of DNA which give at least a 500 1000-fold increase in detection of membrane bound nucleic acid over oilgonucleotide probes. The use of a novel class of these nucleic acid superstructures is described as a specific probe for EBER-1 detection. When two PTLD patients were analyzed for viral load with DNA dendrimers, at least one in 250000 peripheral blood mononuclear cells were shown to be infected with EBV.

Author List

Orentas RJ, Rospkopf SJ, Casper JT, Getts RC, Nilsen TW

Author

James Casper MD Emeritus Professor in the Pediatrics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Cell Line
DNA, Viral
Evaluation Studies as Topic
Genetic Techniques
Herpesvirus 4, Human
Humans
Leukocytes, Mononuclear
Lymphoma
RNA, Viral
Reverse Transcriptase Polymerase Chain Reaction
Transplantation
Viral Load
Virology