OBJECTIVETo evaluate the subgingival prevalence of human cytomegalovirus (HCMV), Epstein-Barr virus-1 (EBV-1) in chronic periodontitis (CP) patients before and after treatment and to analyze the relationship between the prevalent variance and periodontal clinical parameters.

METHODSGingival crevicular fluids of 13 CP patients were collected at baseline, 2 weeks, 2 months and 4 months after periodontal mechanical treatment. HCMV and EBV-1 were detected using nested polymerase chain reaction (n-PCR).

RESULTSThe plaque index (PLI), probing depth (PD) and bleeding index (BI) of CP patients at 2 months, 4 months after periodontal mechanical treatment were evidently lower than before treatment, P < 0.01. These parameters at 4 months after treatment were higher than at 2 months, the differences were significant, P < 0.05. The prevalence of HCMV and EBV in CP patients was 42% (33/78), 14% (11/78). EBV and HCMV were mostly coexistent in the same site [9 sites HCMV(+) in 11 EBV positive sites]. The sites of HCMV(+) and EBV(+) were almost deep pockets. Thirteen of 14 sites with deep pockets were HCMV(+), 9 sites were deep pockets in 11 sites EBV(+). The prevalence of HCMV and EBV (8% and 0 respectively) at 2 weeks was the lowest in all four time points. The prevalence of HCMV and EBV at 2 weeks, 2 months and 4 months following treatment was significantly lower than baseline (P < 0.01), but the prevalence of HCMV (15%) at 2 months after treatment was higher than at 2 weeks (8%), the difference was not significant (P = 0.133).

CONCLUSIONSHerpesviruses may play a role in the development of CP. The changes of the prevalence of herpesviruses before the changes of clinical parameters could be detected after periodontal mechanical treatment. The patients should be re-evaluated and re-treated within 2 months after treatment.

Chronic Periodontitis ; therapy ; Cytomegalovirus ; isolation & purification ; Gingival Crevicular Fluid ; virology ; Herpesvirus 4, Human ; isolation & purification ; Humans

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 investigate human cytomegalovirus (HCMV) and Epstein-Barr virus-type 1 (EBV-1) in GCF and saliva during experimental gingivitis in Chinese young subjects and to evaluate the effect of the virus in the initial stage of gingival inflammation.

METHODSGCF of 14 and 45 and saliva without stimulating in 11 Chinese young males with healthy gingiva were collected at baseline (day 0), day 7, 14 and 21 after stopping oral hygiene and day7 after reestablishing oral hygiene (day 28). DNA of HCMV and EBV-1 were detected by nested-polymerase chain reaction (n-PCR) at the times mentioned above.

RESULTSHCMV was detected in GCF of 4 subjects at baseline, 4 subjects at day 7, 3 subjects at day 14 and 2 subjects at day 21 while the subjects were different. At day 28 HCMV could not be detected. EBV-1 was not detectable in GCF during the experimental gingivitis. HCMV was detected in saliva in 4 subjects and EBV-1 was in 3 subjects. And there is no relationship between the detection of the herpesviruses and the clinical parameters as well.

CONCLUSIONWe suggest that HCMV and EBV-1 are not the important factors during the initial stage of gingival inflammation.

Adult ; Cytomegalovirus ; isolation & purification ; Gingival Crevicular Fluid ; virology ; Gingivitis ; virology ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Young Adult

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*

There has been increasing interest in standardized and quantitative Epstein-Barr virus (EBV) DNA testing for the management of EBV disease. We evaluated the performance of the Real-Q EBV Quantification Kit (BioSewoom, Korea) in whole blood (WB). Nucleic acid extraction and real-time PCR were performed by using the MagNA Pure 96 (Roche Diagnostics, Germany) and 7500 Fast real-time PCR system (Applied Biosystems, USA), respectively. Assay sensitivity, linearity, and conversion factor were determined by using the World Health Organization international standard diluted in EBV-negative WB. We used 81 WB clinical specimens to compare performance of the Real-Q EBV Quantification Kit and artus EBV RG PCR Kit (Qiagen, Germany). The limit of detection (LOD) and limit of quantification (LOQ) for the Real-Q kit were 453 and 750 IU/mL, respectively. The conversion factor from EBV genomic copies to IU was 0.62. The linear range of the assay was from 750 to 10⁶ IU/mL. Viral load values measured with the Real-Q assay were on average 0.54 log₁₀ copies/mL higher than those measured with the artus assay. The Real-Q assay offered good analytical performance for EBV DNA quantification in WB.
DNA, Viral/*blood/metabolism ; Epstein-Barr Virus Infections/diagnosis/virology ; Herpesvirus 4, Human/*genetics/isolation & purification ; Humans ; Limit of Detection ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction