Reverse-transcriptase polymerase chain reaction has been used to analyze the expression of 2 latent genes (EBNA-1 and LMP-1) and one replicative gene (BZLF-1) of Epstein-Barr virus in mononuclear cells from bone marrow and peripheral blood of healthy donors. EBV-gene transcription was detected in 8
Frequent detection of the replicative form of TT virus DNA in peripheral blood mononuclear cells and bone marrow cells in cancer patients
โ Scribed by Sheng Zhong; Winnie Yeo; Mandy Tang; Cuiling Liu; Xiao-rong Lin; Wing M. Ho; Pun Hui; Philip J. Johnson
- Publisher
- John Wiley and Sons
- Year
- 2002
- Tongue
- English
- Weight
- 315 KB
- Volume
- 66
- Category
- Article
- ISSN
- 0146-6615
- DOI
- 10.1002/jmv.2163
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โฆ Synopsis
Abstract
The TT virus (TTV), a member of a family of human viruses related to the circoviridae viruses, was associated initially with acute and chronic liver diseases. TTV consists of a singleโstranded, circular DNA genome of 3.8 kilobases (kb) and at least three open reading frames (ORFs). The objective of the present study was to determine whether or not TTV replicated in peripheral blood mononuclear cells (PBMCs) and bone marrow cells (BMCs). DNA was extracted from the PBMCs or BMCs of 153 cancer patients and from the PBMCs of 50 healthy blood donors (the controls). By using a single round of polymerase chain reaction (PCR), TTV was detected in 98.6% (141 of 143) of the PBMCs and in 90% (9 of 10) of the BMCs from cancer patients. TTV DNA was detected in significantly fewer control subjects at 86% (43 of 50; Pโ<โ0.05). Strandโspecific PCR (SSPCR) targeting the ORF2 of the common genotypes of TTV was developed specifically to detect TTV positive or negative strand DNA and to examine TTV replication. TTV positive strand DNA, which may be an intermediate of viral replication, was detected in 55.3% (78 of 141) of the TTVโinfected PBMCs of the cancer patients and in 7% (3 of 43) of the controls (Pโ<โ0.001). The replicative form of TTV was also detectable in 55.6% (5 of 9) of the TTVโinfected BMCs. The existence of doubleโstrand (positive and negative strands) TTV DNA in PBMCs and BMCs of the cancer patients was also supported by the finding that TTV DNA extracted from these cells was resistant to S1 nuclease. Using in situ hybridization, TTV DNA was also demonstrated to be present in the nucleus of PBMCs. It is concluded that replicative intermediate forms of TTV DNA are present in both PBMCs and BMCs, indicating that blood cells may be a site of TTV replication. J. Med. Virol. 66:428โ434, 2002. ยฉ 2002 WileyโLiss, Inc.
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