To analyze the association of Epstein-Barr virus (EBV) with gastrointestinal non-Hodgkin's lymphomas arising in immunocompetent patients, 56 consecutive cases of gastrointestinal lymphomas (B-cell: 52-cases, T-cell: 3 cases, T/NK-cell: 1 case) occurring in the stomach (33 cases), intestine (22 cases) and esophagus (1 case) were investigated for the presence of EBV using polymerase chain reaction analysis as a screening method followed by EBER-1 RNA and DNA in situ hybridization (ISH) and immunohistochemistry for the expression of latent membrane protein 1 (LMP-1). Forty-seven cases demonstrated extractable DNA and EBV DNA was detected only in 4 cases. Among the, RNA (EBER-1) and DNA ISH analysis confirmed the presence of the EBV genome in tumor cells in 3 cases (T/NK-cell lymphoma of ileum, gastric high-grade B-cell lymphoma of mucosa-associated lymphoid tissue, gastric diffuse large B-cell lymphoma). Only the T/NK cell lymphoma showed diffuse positivity of tumor cells while 2 gastric B-cell lymphomas demonstrated a scattered positive reaction and no cases expressed LMP-1. Nine cases without extractable DNA by the PCR method showed no nuclear signal by EBER-1 ISH. These findings suggest that most sporadic primary gastrointestinal lymphomas in Korea are not associated with EBV.
Gastrointestinal Neoplasms/virology* ; Gastrointestinal Neoplasms/pathology ; Genome, Viral ; Herpesvirus 4, Human/isolation & purification* ; Herpesvirus 4, Human/genetics ; Human ; In Situ Hybridization ; Korea ; Lymphoma/virology* ; Lymphoma/pathology ; Polymerase Chain Reaction

The development of a multiplex polymerase chain reaction (PCR) method for rapid and accurate detection and typing of herpes simplex virus type 1 (HSV-1), and type-2 (HSV-2), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) is very important for clinical diagnosis to allow the deliver of therapy as early as possible. Large scale amplifications by multiplex PCR of viral DNA can lower the cost and time for viral diagnosis. In this study, therefore sensitive quadruplex PCR was achieved by optimizing parameters such as primers, and 1.5 mM magnesium and 200 uM dNTPs concentrations. The concentrations of HSV-1, HSV-2, CMV and EBV primers were 0.5, 0.3, 0.25 and 0.25 pmoles, respectively. Optimal annealing temperature was 54 degrees C. Employing these conditions, we could detect 10 copies of reconstructed template plasmid DNA, which were cloned to vectors containing target sequences of viral DNA. PCR products of 271 bp for HSV-1, 231 bp for HSV-2, 368 bp for CMV, and 326 bp for EBV were separated on 5.0% polyacrylamide gel electrophoresis and confirmed by direct sequencing. The present study showed that the quadruplex PCR assay described herein has potential application in clinical diagnosis, when rapid, accurate detection and typing of viruses HSV-1, HSV-2, CMV or EBV are necessary.
Cytomegalovirus/classification/*isolation & purification ; Herpesvirus 1, Human/classification/*isolation & purification ; Herpesvirus 2, Human/classification/*isolation & purification ; Herpesvirus 4, Human/classification/*isolation & purification ; Human ; *Polymerase Chain Reaction/methods ; Sensitivity and Specificity ; Support, Non-U.S. Gov't

BACKGROUNDThe dilemma of pathogens identification in patients with unidentified clinical symptoms such as fever of unknown origin exists, which not only poses a challenge to both the diagnostic and therapeutic process by itself, but also to expert physicians.

METHODSIn this report, we have attempted to increase the awareness of unidentified pathogens by developing a method to investigate hitherto unidentified infectious pathogens based on unbiased high-throughput sequencing.

RESULTSOur observations show that this method supplements current diagnostic technology that predominantly relies on information derived five cases from the intensive care unit. This methodological approach detects viruses and corrects the incidence of false positive detection rates of pathogens in a much shorter period. Through our method is followed by polymerase chain reaction validation, we could identify infection with Epstein-Barr virus, and in another case, we could identify infection with Streptococcus viridians based on the culture, which was false positive.

CONCLUSIONSThis technology is a promising approach to revolutionize rapid diagnosis of infectious pathogens and to guide therapy that might result in the improvement of personalized medicine.

Female ; Herpesvirus 4, Human ; genetics ; isolation & purification ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Male ; Viridans Streptococci ; genetics ; isolation & purification

Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Infectious Mononucleosis ; drug therapy ; pathology ; virology ; RNA, Viral ; isolation & purification ; Sensitivity and Specificity

OBJECTIVETo compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC.

METHODSThe plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted.

RESULTSEBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion >/= 1:40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA >/= 1:40 test (0.4839, 0.5385 and 1.0480, respectively).

CONCLUSIONThe EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC.

Antigens, Viral ; blood ; DNA, Viral ; blood ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunoglobulin A ; blood ; Nasopharyngeal Neoplasms ; diagnosis ; virology ; Serologic Tests

Child ; Epstein-Barr Virus Infections ; complications ; Female ; Giardia lamblia ; isolation & purification ; Giardiasis ; complications ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Lymphohistiocytosis, Hemophagocytic ; drug therapy ; etiology

The precise etiology of hemolytic uremic syndrome (HUS) is unknown. However, it has been associated with bacterial (Shigella, Salmonella, E. coli, S. pneumoniae), Bartonella, and viral (coxsackie, ECHO, influenza, varicella. Epstein-Barr) infections and with endotoxemia. Recently, we experienced a case of HUS in a 16-year-old boy who was in the acute phase of an Epstein-Barr virus (EBV) infection. He had typical manifestations of HUS and EBV infection. He also transiently presented disseminated intravascular coagulation. His renal dysfunction recovered by supportive care, including hemodialysis, plasmapheresis, antihypertensive medication and aspirin. We present this case with a review of the literature as the second report of HUS associated with EBV infection.
Adolescence ; Follow-Up Studies ; Hemolytic-Uremic Syndrome/virology* ; Hemolytic-Uremic Syndrome/therapy ; Herpesviridae Infections/diagnosis* ; Herpesvirus 4, Human/isolation & purification* ; Human ; Male ; Renal Dialysis ; Tumor Virus Infections/diagnosis*

OBJECTIVETo prepare an Epstein-Barr virus (EBV) microarray using known and predicted EBV-coded genes as the cDNA probes to detect the EBV gene expression in nasopharyngeal carcinoma (NPC) tissues.

METHODSThe EBV gene probes were amplified by PCR using a pair of primers designed in both sides of the multiple clone site (MCS) of the T/A vector. After purification of the PCR products, 85 EBV genes and 8000 human genes were printed onto the same slide as the detection chip consisting of both EBV and human genes. This genechip was used to detect the differential gene expression in NPC and non-cancerous nasopharynx (NP) tissues.

RESULTSDetection of the human gene expression profile using the prepared genechip resulted in the identification of numerous human genes in the tissue specimens. Some EBV genes were also detected in the tissues using the genechip, but the signals of the genes appeared rather weak without distinctly visible fluorescence, and were not comparable to the strong signal intensities of the human genes.

CONCLUSIONThe EBV microarray, though constructed successfully, can not meet the needs for clinical application due to the limited detection sensitivity and the relative small quantity of EBV gene expression in NPC samples. Further improvements of the research methods are warranted.

Carcinoma, Squamous Cell ; virology ; Gene Expression Profiling ; Genome, Viral ; genetics ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Nasopharyngeal Neoplasms ; virology ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo investigate the relationship between Epstein-Barr virus (EBV) infection and chronic periodontitis.

METHODSIn 62 chronic periodontitis patients, subgingival plaque samples were collected from active periodontitis site, quiescent periodontitis site, and initial gingivitis sites of each subject. The nested polymerase chain reaction (nested-PCR) was used to determine the presence of EBV.

RESULTSThe EBV was detected in 58.0% of the samples from active periodontitis sites, which was significantly higher than that from quiescent periodontitis sites (58.0% vs 22.6%, P < 0.01) and from initial gingivitis sites (58.0% vs 19.4%, P < 0.01).

CONCLUSIONThe results suggest that EBV infection is associated with the activity of chronic periodontitis.

Adult ; Chronic Disease ; Epstein-Barr Virus Infections ; complications ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Periodontitis ; virology ; Polymerase Chain Reaction

A CD30+ anaplastic large cell lymphoma (ALCL) cell line was established from the mononuclear cells isolated from pleural effusion of a patient with non-Hodgkin's lymphoma. The cell line's biological characteristics were analyzed. The results showed that the established cell line could survive and proliferate in RPIM 1640 medium; the Wright-Giemsa-stained cells were exactly similar to malignant cells of CD30+ ALCL in morphology, with many diffuse virus granules in cytoplasm; the cytochemical staining of the cells showed the following reactivity pattern: positive for acid phosphatase (ACP) and periodic acid-Schiff (PAS), negative for peroxidase (POX), myeloperoxidase (MPO) and platelet peroxidase (PPO). The immunoprofile of the cells was positive for CD45, HLA-DR, CD30 and negative for EMA, CD34, CD38, CD2, CD3, CD4, CD7, CD8, CD10, CD15, CD19 and CD20. The cytogenetic analysis showed complicate d qualitative and quantitative abnormality of chromosomes, without typical t(2;5). It is concluded that the established cell line is CD30+ anaplastic large cell lymphoma cell line.
Apoptosis ; Cell Line, Tumor ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunophenotyping ; Karyotyping ; Ki-1 Antigen ; analysis ; Lymphoma, Large B-Cell, Diffuse ; genetics ; immunology ; pathology ; Middle Aged