Epstein-Barr Virus Polymerase Chain Reaction Research Articles

Post-transplant lymphoproliferative disorder associated Epstein-Barr virus DNAemia after liver transplantation in children: Experience from single center.

The most prevalent malignancy that complicates both adult and pediatric solid organ transplantation is post-transplant lymphoproliferative disorder (PTLD). This study aimed to analyze the clinical and pathological characteristics, treatments, and outcomes of Epstein-Barr virus (EBV) DNAemia and PTLD in pediatric liver transplant recipients. A retrospective chart review was performed on 112 patients less than 18 years of age who underwent isolated orthotopic liver transplantation (OLT) between 2010 and 2022 at Ege University Children's Hospital. Data gathered for 1-year post-OLT included age at OLT, EBV, immunoglobulin (Ig)M/IgG status of the donor and recipient, indication for OLT, induction regimen, all immunosuppression levels, date and result of EBV polymerase chain reaction testing, rejection episodes documented by liver biopsy, and the development of PTLD. Forty-nine patients (43.75%) developed EBV DNAemia (median interval from surgery: 2 months, min-max: 2-36), of which 43 (87.8%) grafts came from living donors, and 6 (12.2%) came from deceased donors. Nine (18.4%) patients died during follow-up, and eight (16.3%) developed PTLD. Of these 8 patients; five patients developed EBV-related disease, one child developed hemophagocytic lymphohistiocytosis, one developed aplastic anemia, and one child developed B cell lymphoma. When PTLD patients and without-PTLD patients were compared, pediatric intensive care unit hospitalization, abnormal bone marrow biopsy findings, lymphadenopathy, age at diagnosis of EBV DNAemia, EBV viral load, tacrolimus (FK 506) pre-infection, were higher and tacrolimus 1-month levels were lower in patients with PTLD (p < 0.05). In logistic regression analysis, we showed that the age at diagnosis of EBV DNAemia was significantly higher in children with PTLD (p = 0.045; OR: 1.389; 95% CI: 1.007-1.914). PTLD is a rare but severe complication associated with EBV after OLT. This study demonstrated that PTLD is associated with older age, higher tacrolimus blood levels before EBV DNAemia, and higher peak EBV viral load at 1 month of EBV DNAemia.

Risk factors for persistent fever in children with infectious mononucleosis.

Fever is a common symptom in children with infectious mononucleosis (IM). However, the significance of fever duration is poorly understood. This study aimed to analyze the risk factors for persistent fever in children with IM. Patients diagnosed with IM (aged <18 years; except those with concomitant hematological malignancies or tumor diseases) in a high-volume academic hospital in 2019 were reviewed from a prospectively maintained database. Comparison analysis was conducted between children with transient fever (≤7 days) and those with persistent fever (>7 days). The risk factors for persistent fever in children with IM were examined using binary logistic regression. Furthermore, the predictive ability of these risk factors was assessed and presented using a receiver operating characteristic (ROC) curve. Of the 184 children included in this study, 131 (71.96%) belonged to the transient fever group and 53 (28.04%) belonged to the persistent fever group (median age: 49 and 64 months, respectively; P = 0.093). Statistical significance was observed in hepatomegaly, alanine aminotransferase level, blood triglyceride level, and blood Epstein-Barr virus polymerase chain reaction (EBV-PCR) copy number (all P < 0.05). Binary logistic regression revealed that high blood triglyceride level was the risk factor for persistent fever in children with IM. High blood triglyceride level predicted persistent fever duration, with an area under the ROC curve of 0.73 and an optimal cutoff value of 1.315 mmol/L. High blood triglyceride level was the risk factor for persistent fever in children with IM. Thus, children with elevated levels of blood triglycerides need additional care. To diagnose IM, blood EBV-PCR is more useful than throat swab EBV-PCR.

EBV-POSITIVE B-CELL ULCERATIVE PROLIFERATION IN THE ORAL CAVITY

<h3>Objective</h3> Lymphoproliferative disorders (LPDs) are a group of conditions characterized by excessive production of lymphocytes manifested by lymphadenopathy, monoclonal lymphocytosis, and bone marrow infiltration. LPDs are often associated with Epstein-Barr virus (EBV) infection. EBV-associated LPDs most commonly appear in the setting of immunocompromised status such as HIV and in transplantation with the extended use of immunosuppressive medications. Individuals with primary immunodeficiency disorders, such as common variable immunodeficiency disease (CVID) or severe combined immunodeficiency, are also at risk of LPD. <h3>Case Summary</h3> A 67-year-old woman complained of mouth ulcerations for 3 months before presentation that were progressively getting worse. Past medical history included CVID, CD4 lymphopenia, rhinosinusitis, eosinophilic asthma, chronic back pain, anxiety, and depression. Medications included immunoglobulin replacement therapy, mepolizumab, montelukast, albuterol, cetirizine, bupropion, clonazepam, fluoxetine, and gabapentin. The patient was allergic to penicillin and azithromycin. Past surgical history was notable for removal of skin melanoma and total knee replacement. Review of systems included a 3-month, 25-lb weight loss, odynophagia, constipation, weakness, and fatigue. Extraoral examination was unremarkable. Intraoral examination revealed 3 × 2 cm² necrotizing palatal ulcerations bilaterally. Necrotizing ulceration was also noted on multiple areas of the gingiva. Differential diagnosis included acute necrotizing ulcerative periodontitis, deep fungal infection (aspergillosis), and lymphoma/LPD. Complete blood count with differential, EBV polymerase chain reaction (PCR), cytomegalovirus PCR and maxillofacial computed tomography (CT) were ordered. The CT scan was unremarkable and laboratory evaluation was negative except for positive EBV PCR. Oral biopsy demonstrated EBV positive B-cell ulcerative proliferation. Positron emission tomography/CT showed widespread fluorodeoxyglucose (FDG)-avid lymphadenopathy. An axillary lymph node biopsy showed atypical lymphoid proliferation in a background of atypical and reactive follicular hyperplasia most consistent with EBV-negative follicular lymphoma, which appeared histologically distinct from the oral EBV-positive LPD. The patient is currently being treated with single-agent rituximab therapy. <h3>Conclusions</h3> EBV may cause oral diseases including infectious mononucleosis, oral hairy leukoplakia, necrotizing ulcerative mucositis, non-Hodgkin's lymphoma, and EBV-related LPD. Persistent oral lesions in patients with CVID could be related to underlying systemic disease. This case is unique because the patient seemingly had follicular lymphoma unrelated to her EBV-positive LPD oral disease, yet in the setting of severe immunosuppression from CVID.

Hepatosplenomegaly and Periventricular Cyst in a Neonate with Direct Hyperbilirubinemia

Hepatosplenomegaly and Periventricular Cyst in a Neonate with Direct Hyperbilirubinemia

Association of HPV and EBV in Oral Verrucous Squamous Cell Carcinoma and Oral Verrucous Hyperplasia

Objective Oral verrucous squamous cell carcinoma or oral verrucous carcinoma (OVC) is a rare verrucous variant of oral squamous cell carcinoma (OSCC), which accounts for 2 to 12% of all oral carcinomas. Oral verrucous hyperplasia (OVH) is clinically similar to OVC and has been proposed to be a precursor lesion of OVC. Etiopathogenesis of both lesions is still inconspicuous. Oncogenic viruses such as human papillomavirus (HPV) and Epstein–Barr virus (EBV) have been reported to be associated with some cases of OSCC, and we hypothesized that it may act as a causative agent of these verrucous lesions. This study aimed to investigate frequency of HPV and EBV infections in OVC and OVH. Material and Methods Using polymerase chain reaction (PCR), a total of 35 formalin-fixed paraffin-embedded (FFPE) tissue samples, including 27 OVC samples and 8 OVH samples, were investigated for HPV and EBV infection. HeLa and B95-8 cell lines were used as positive controls of HPV and EBV PCR, respectively. Results All OVC and OVH samples show a positivity to GAPDH , whereas neither HPV nor EBV PCR products was detected in both OVC and OVH samples. Conclusions In summary, our study demonstrated that HPV and EBV are not involved in pathogenesis of OVC and OVH. Other etiologic factors contributing to OVC and OVH need to be further clarified.

Clinical significance of Epstein-Barr virus polymerase chain reaction in cerebrospinal fluid.

Epstein-Barr virus (EBV) is implicated in various neurological conditions. However, the relationship between EBV DNA in cerebrospinal fluid (CSF) and central nervous system (CNS) infection is unclear. We evaluated the clinical manifestation of patients with EBV DNA detected in CSF. We reviewed the medical records of patients admitted to Seoul National University Hospital from January 2000 to March 2021 who underwent EBV polymerase chain reaction (PCR) tests in CSF. The subjects were divided into positive and negative groups depending on the presence of EBV DNA, and further clinical information was obtained from positive patients. CSF EBV PCR tests were performed in 807 patients, and 57 (7.1%) tested positive. Pleocytosis was common (81.1%) in CSF samples with EBV DNA detected, and the proportion was significantly higher than that in samples that were EBV PCR negative (44.5%, p < 0.0001). Among 57 patients with EBV DNA detected in CSF, 51 (89.5%) were diagnosed with CNS infection or inflammatory disorders. Of the 51 patients, 31 (60.8%) had possible etiologies other than EBV. Follow-up evaluation was conducted in 19 of 20 patients, and 63.2% showed a favorable outcome. Positive EBV PCR in CSF is mostly nonspecific and should be interpreted with caution. A comprehensive workup is needed to identify other etiologies before considering EBV as the sole culprit.

Lues Maligna in an Immune-Reconstituted Patient with Retroviral Infection: A Case Report

Lues Maligna in an Immune-Reconstituted Patient with Retroviral Infection: A Case Report

Simultaneous Assessment of Plasma and Peripheral Mononuclear Cells for Multiple Viral Load Quantification in Peripheral Blood of Patients after Heart Transplantation

Purpose Epstein-Barr Virus (EBV) is well known to be associated with posttransplant lymphoproliferative disorders (PTLD). Due to moderate immunosuppression in heart transplant (HTx) recipients, many viruses other than EBV are also infected and activated and assessment of simultaneous viral loads of other viruses is also important. In the present study, the role of simultaneous assessment of plasma and peripheral mononuclear cells for multiple viral load quantification in peripheral blood of pediatric patients after heart transplantation was elucidated. Methods In a period of 9 months (from January to October 2009) we collected 28 samples of 3 adult and 6 pediatric cardiac transplant recipients. Quantitative polymerase chain reaction for EBV, Cytomegalovirus, Parvovirus 19 (B19), BK and JC virus, and HHV6 was simultaneously performed based on the TaqMan system using DNA which isolated from peripheral mononuclear cells (PBMCs) and plasma, as described elsewhere. Phenotyping of infected lymphocytes were also done. CD8, CD19, CD45RO, CD57, etc. Results Case 1: 2-year old girl who underwent HTx at 1 year of her age, developed B cell type PTLD and died of cytokine release syndrome, showed rapid increases in EBV load in both PMBC and plasma as well as HHV 6 and B19. Case 2: 22-year female with subclinical infection for 19 years after HTx, showed sudden decrease in both in EBV load in both PMBC and plasma after conversion of everolimus to mycophenolate mofetil and both EBV loads eliminated after cytotoxic T lymphocyte (CTL) formation against EBV. Three patients (clinical in 2 and subclinical in 1) showed an increase in B19 load. Although B19 load became negative in 2 patients, B19 load sustained positive for 9 months in one patients who experienced B cell type PLTD 6 years ago and had continuous B19 infection. In one 38-year old male with steroid treatment for nephrosis and one 11-year old boy with living renal transplantation 10 years after HTx showed increases in BK and JC virus loads and an rapid decrease in both viral loads after reduction of immunosuppression. Conclusion Simultaneous multiple viral load quantifications are useful to evaluate immunosuppressive status of HTx recipients as well as monitoring of viral infection itself.

Risk of Post-Transplant Lymphoproliferative Disorders (PTLD) in Solid Organ Transplantation Patients with EBV Viremia

Introduction Post-transplant lymphoproliferative disorders (PTLD) are mostly Epstein-Barr virus (EBV) positive lymphoid proliferations resulting from immunosuppression following allogeneic stem cell or solid organ transplantation (SOT). Despite this strong association, there is a lack of data evaluating the risk of PTLD in SOT patients (pts) with (w/) EBV viremia. The significance of EBV detection remains unclear and can lead to unwarranted preemptive treatments in SOT pts. The aim of this study is to assess the association between peripheral blood EBV viral load and risk of PTLD post SOT. Methods We identified 6468 adult and pediatric pts from the Cleveland Clinic SOT database who underwent SOT from 2002-2016. We included pts who had ≥1 EBV viral load test by PCR, and excluded pts w/ first EBV test at or after PTLD diagnosis. EBV viral load was quantified by polymerase chain reaction (PCR) of whole blood samples and expressed as log copies/mL. Lower detection limits varied from 2.00 to 2.70 log copies/mL. EBV monitoring post SOT was done at the physician9s discretion. Potential risk factors for PTLD were assessed using Fine and Gray competing risk regression. Stepwise analysis was used to identify multivariable risk factors. Landmark analyses at 2 and 6 months post SOT (LM-2 and LM-6) were performed based on pts9 peak pre-landmark EBV levels. Pts were excluded if they died, lost follow-up (f/u) or developed PTLD prior to the landmark. Two multivariable models were assessed at each landmark using EBV level cutoffs of 3.00 and 4.00 log. Results Of 6324 pts w/ available f/u data, 3348 (52%) had ≥1 quantitative EBV PCR test and were included in this analysis w/ a total of 91,067 tests performed. Median age at SOT was 54 years (range 0.3-79) and 63% were male. The most common SOT was kidney (38%) followed by lung (37%), liver (18%), heart (7%), pancreas (6%) and intestine (2%), w/ 7% receiving >1 organ type (Table 1). Of 3348 pts included, 1503 (45%) developed EBV viremia and 68 (2%) developed PTLD; 45/1503 (3.0%) and 23/1845 (1.2%) of pts w/ and without (w/o) EBV viremia, respectively. EBV was detected in 44% of pts w/o PTLD and 66% of pts w/ PTLD. Median age at PTLD diagnosis was 58 years (range 1-75), with a median time from SOT to PTLD of 33 months (range 2-146). PTLD morphology was DLBCL (n=39), polymorphic (n=9) Burkitt (n=5), other (n=10) and unknown (n=5). EBV-encoded RNA (EBER) testing in tumor was positive in 47%, negative in 44% and unknown in 9% of cases. In pts w/ and w/o PTLD, tacrolimus was the most common immunosuppressant used at 1 month (99% and 89%), 1 year (94% and 88%) and 2 years (81% and 74%) post SOT, respectively. First EBV was checked at a median of 54 days (range 0-5090) in pts w/o PTLD and 28 days (range 0-4308) in pts w/ PTLD. The median number of EBV tests/pt before PTLD or last f/u was 4 (range 1-214) in pts w/o PTLD and 6 (range 1-123) in pts w/ PTLD. Peak EBV levels ≥3.00 and ≥4.00 log were detected in 36% and 13% of pts w/o PTLD, and 60% and 40% of pts w/ PTLD, respectively. Median f/u in both cohorts for pts who were alive was 6.1 years. In multivariable analysis (MVA) for non-EBV risk factors for PTLD, liver (HR 2.19, 95% CI 1.31 - 3.66, p=.003) and intestine SOT (HR 4.62, 95% CI 1.84 - 11.6, p=.001) were the only factors associated w/ higher PTLD risk; age, years since SOT, gender, race, other types of SOT and >1 SOT were not significant. 1685 pts were eligible for LM-2 (43 w/ PTLD) and 2093 for LM-6 (36 w/ PTLD). In univariable analysis at LM-2 and LM-6, peak EBV levels ≥4.00 log (LM-2: HR 2.72, p=.016; LM-6: HR 7.08, p Conclusion In this large cohort of SOT pts, Disclosures Hill:Pharmacyclics: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Novartis: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Genentech: Honoraria, Membership on an entity9s Board of Directors or advisory committees, Research Funding; Abbvie: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Seattle Genetics: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Abbvie: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Amgen: Research Funding; Pharmacyclics: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Novartis: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Seattle Genetics: Honoraria, Membership on an entity9s Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity9s Board of Directors or advisory committees.

Epstein-Barr virus infection in children with renal transplantation: A single-centre experience.

This study aimed to investigate the incidence, timing, manifestations, managements, and outcomes of Epstein-Barr virus (EBV) infection in paediatric renal transplant recipients in Korea. We retrospectively evaluated 70 patients aged <18 years who had undergone renal transplantation between January 1990 and November 2014 at a single centre in Korea. EBV infection was diagnosed via serological test or real-time quantitative polymerase chain reaction (PCR). The diagnosis of post-transplant lymphoproliferative disorder (PTLD) was based on biopsy findings. In total, 21 patients (30.0% of renal transplant recipients) had EBV infection. EBV infection occurred at an average age of 12.6 ± 4.5 (median, 12.0; range, 7.0-24.0 years, with a mean period of 28.3 ± 27.2 (median, 14.0; range, 2.0-75.0) months for developing EBV infection after transplantation. EBV infection developed 12 times more frequently in pre-transplant EBV-seronegative recipients. Eight patients (38% of EBV-infected patients) had EBV disease, and six patients (75% of patients with EBV disease) had PTLD. The maximum EBV PCR titer was greater in patients with EBV disease than in the asymptomatic EBV infection group. The main treatment for EBV infection was the reduction in immunosuppressants. Asymptomatic EBV infection resolved in approximately 80% of the patients. One patient (17% of the patients with PTLD) expired. The glomerular filtration rate did not deteriorate during the treatment of EBV infection. Regular EBV monitoring in renal transplant recipients is mandatory for early diagnosis and treatment of EBV infections and prevention of PTLD, especially in pre-transplant EBV IgG-negative patients.

Epstein–Barr Virus DNAemia and co-occurrence with cytomegalovirus DNAemia in postrenal transplant recipients from a tertiary care center

Aim: Co-occurrence of Epstein–Barr virus (EBV) with Cytomegalovirus (CMV) is associated with an increased risk of EBV-associated posttransplant lymphoproliferative disorder (PTLD). Quantitation of EBV by real-time polymerase chain reaction (PCR) can aid the clinicians in the initiation of preemptive measures to improve the survival of the graft. Methods: The study was conducted among postrenal transplant recipients (PRTRs) who were attending the nephrology department from 2011 to 2016. Real-time quantitative PCR for EBV was performed in whole blood. PRTRs were classified into asymptomatic with altered renal parameters (Group A) and symptomatic (Group B), which were further subcategorized into Group B1 (fever with anemia, leukopenia, thrombocytopenia, or altered liver enzymes (any two), Group B2 (Group B1 + end-organ disease or only end-organ disease), and Group B3 (graft dysfunction [GDF]). The posttransplant period was also defined. DNA was extracted (Qiagen, Hilden, Germany) from whole blood, and real-time PCR was performed using QuantiTect multiplex PCR kit. Unpaired t-tests and ANOVA were used to analyze the data. Results: A total of 89 PRTRs were enrolled, of which 39.3% (n = 35) had EBV DNAemia, 43.1% during very late, 41.1% in late and 28.6% in immediate post transplant periods. EBV DNAemia ranged from 324 to 32,436 copies/ml. EBV DNAemia was found in 84% (n = 75) of symptomatic (Group B) and 16% (n = 14) of asymptomatic (Group A). Among the PRTRs with GDF (Group B3), 44% (n = 11/25) had EBV DNAemia of 2893.9 ± 1869 copies/ml. EBV DNAemia was considerably higher in PRTRs without GDF (8700.2 ± 9675.6 copies/ml) than PRTRs with GDF and the difference was statistically significant (P = 0.004). EBV DNAemia with CMV DNAemia among PRTRs was found in 21.3% (n = 19). Conclusion: High EBV DNAemia may precede PTLD or GDF; therefore, regular screening of EBV DNAemia is warranted. CMV and EBV DNAemia may also co-exist in PRTRs. As CMV is an immunomodulating virus, it increases the risk of opportunistic infections, especially EBV.

Polymerase Chain Reaction and Goldmann-Witmer Coefficient to Examine the Role of Epstein-Barr Virus in Uveitis

ABSTRACTPurpose: To use polymerase chain reaction (PCR) and Goldmann-Witmer Coefficient (GWC) calculation to search for evidence that Epstein-Barr virus (EBV) causes uveitis.Methods: A prospective cross-sectional study where participants with positive multiplex EBV PCR results were further investigated by: 1) real-time PCR for EBV viral loads (VL) and 2) EBV GWC.Results: Eleven of 106 consecutive uveitis patients (10.4%) had positive multiplex PCR for EBV on aqueous humor sampling and 7/11 (63.6%) were HIV-positive. Only 4/10 (40%) cases had detectable intraocular EBV VLs which were always lower than the blood or plasma VL. EBV GWC was negative in all 10 cases tested. In 9/11 (81.8%) of these cases an alternative, more plausible cause of uveitis was identified.Conclusion: We found no evidence of active intraocular replication or antibody production to prove that EBV caused uveitis in these cases. In most cases an alternative treatable cause of uveitis was identified.

Paralysie faciale bilatérale au cours d’une infection à virus d’Epstein–Barr

Bilateral facial nerve palsy is a rare and sometimes difficult diagnosis. We describe a case of bilateral simultaneous facial nerve palsy associated with Epstein-Barr virus (EBV) infection in a 3-year-old boy. Several symptoms led to the diagnosis of EBV infection: the clinical situation (fever, stomachache, and throat infection), white blood cell count (5300/mm3 with 70% lymphocyte count), seroconversion with EBV-specific antibodies, lymphocytic meningitis, and a positive blood EBV polymerase chain reaction (9.3×103 copies of EBV-DNA). An MRI brain scan showed bilateral gadolinium enhancement of the facial nerve. A treatment plan with IV antibiotics (ceftriaxone) and corticosteroids was implemented. Antibiotics were stopped after the diagnosis of Lyme disease was ruled out. The patient's facial weakness improved within a few weeks. Bilateral facial nerve palsy is rare and, unlike unilateral facial palsy, it is idiopathic in only 20% of cases. Therefore, it requires further investigation and examination to search for the underlying etiology. Lyme disease is the first infectious disease that should be considered in children, especially in endemic areas. An antibiotic treatment effective against Borrelia burgdorferi should be set up until the diagnosis is negated or confirmed. Further examination should include a blood test (such as immunologic testing, and serologic testing for viruses and bacterium with neurological tropism), a cerebrospinal fluid test, and an MRI brain scan to exclude any serious or curable underlying etiology. Facial bilateral nerve palsy associated with EBV is rarely described in children. Neurological complications have been reported in 7% of all EBV infections. The facial nerve is the most frequently affected of all cranial nerves. Facial palsy described in EBV infections is bilateral in 35% of all cases. The physiopathology is currently unknown. Prognosis is good most of the time.

Primary Epstein-Barr virus infection, seroconversion, and post-transplant lymphoproliferative disorder in seronegative renal allograft recipients: a prospective cohort study.

Epstein-Barr virus (EBV)-seronegative renal transplant recipients are at risk of post-transplant lymphoproliferative disorder (PTLD). We compared primary EBV infection, seroconversion, and PTLD in EBV-seronegative patients who received renal allograft from seropositive or seronegative donors (D+/R- and D-/R-, respectively). We prospectively followed 25 D+/R- and 8 D-/R- recipients. We followed patients from January 1999 to June 2009 with clinical visits, monthly EBV polymerase chain reaction tests, and serologic tests for a period of 1 year after kidney transplantation and on an individual basis thereafter. Three patients (9%) developed PTLD including 2 early-onset (<12 months) and 1 late-onset (>12 months) disease. In D+/R- and D-/R- patients, the frequencies of PTLD (8% vs. 12.5%, P = 0.7), EBV seroconversion (64% vs. 50%, P = 0.4), and EBV viremia (40% vs. 25%, P = 0.6) were not significantly different. Clinical, serologic, and virologic surveillance as well as reduction in immunosuppression after evidence of primary EBV infection resulted in a PTLD rate of 9%, despite a seroconversion rate of 60.6%. Rate of graft loss after reduction in immunosuppression was 10% (2 of 20), which was not significantly different from 13 patients without EBV seroconversion (no graft loss, P = 0.5). Rates of viremia, seroconversion, and PTLD in D+/R- and D-/R- patients appear to be similar. The incidence of PTLD in renal transplants ranges from 0.5% to 2.9%. Our data show a significantly higher rate in EBV-seronegative renal allograft recipients, suggesting the need for close surveillance. Our data also suggest that donors for EBV-seronegative recipients may be accepted irrespective of positive or negative serostatus, with ongoing surveillance important in either circumstance.

Unusual Indolent Course of a Chronic Active Epstein-Barr Virus-Associated Natural Killer Cell Lymphoproliferative Disorder.

Natural killer (NK) cell lymphoproliferative disorders are uncommon and the Epstein-Barr virus (EBV) plays an important aetiological role in their pathogenesis. We report a 20-year-old male with a chronic active EBV infection associated with a NK cell lymphoproliferative disorder which had an unusual indolent course. He presented to the Sultan Qaboos University Hospital in Muscat, Oman, in December 2011 with a history of intermittent fever and coughing. Examinations revealed generalised lymphadenopathy, hepatosplenomegaly, leukocytosis, transaminitis, diffuse bilateral lung infiltrates and bone marrow lymphocyte involvement. A polymerase chain reaction (PCR) test revealed a high EBV viral load in the peripheral blood cells. The patient received a course of piperacillin-tazobactam for Klebsiella pneumoniae, but no active treatment for the lymphoproliferative disorder. However, his lymphocyte count, serum lactate dehydrogenase and liver enzymes dropped spontaneously. In addition, EBV PCR copies fluctuated and then decreased significantly. He remained clinically asymptomatic over the following four years.

Clinical Utility of Thallium-201 Single Photon Emission Computed Tomography and Cerebrospinal Fluid Epstein-Barr Virus Detection Using Polymerase Chain Reaction in the Diagnosis of AIDS-Related Primary Central Nervous System Lymphoma.

Objective: To determine the diagnostic efficiency of thallium-201 single photon emission computed tomography (SPECT) and Epstein-Barr virus (EBV) polymerase chain reaction (PCR) in the differentiation of primary central nervous system lymphoma (PCNSL) from other central nervous system processes in patients with HIV/AIDS.Design/Methods: Over 10 years, 68 thallium-201 SPECT scans were performed on neurologically symptomatic HIV+ patients with focal lesions on CT or MRI at the Johns Hopkins Hospital. Diagnoses were then established by either autopsy, biopsy, or clinical response to anti-toxoplasmosis therapy. Patients were categorized prior to a prospective clinical reading of the SPECT scans by nuclear medicine physicians.Results: In our patient sample overall, the diagnostic efficiency of thallium-201 SPECT was 79%. The diagnostic accuracy of EBV PCR testing alone in a subset of 22 patients in our study that had CSF analyzed was 73%. However, when both positive EBV PCR and positive thallium-201 SPECT results were used together, the diagnostic accuracy improved to 100% based on a sample of 13 patients where EBV PCR and SPECT imaging results were concordant. Conclusion: Thallium-201 SPECT has a relatively high positive predictive value with regards to the diagnosis of PCNSL, which suggests that patients with positive results could undergo empiric radiation treatment without resorting to brain biopsy. However, the predictive value can be increased by testing for CSF EBV using PCR. Alternatively, if CSF cannot be safely obtained because of mass effect, we believe that these data still suggest that empiric radiation treatment should be considered when discussing treatment options with patients with a positive thallium-201 SPECT.

Severe leptospirosis complicated by Epstein–Barr Virus reactivation

Weil's disease is a severe, potentially fatal illness following Leptospira interrogans infection. The reported case of a patient suffering from acute renal failure, jaundice, thrombocytopenia, rhabdomyolysis and encephalitis syndrome highlights the clinical challenge in reference to Weil syndrome complicated by Epstein-Barr Virus (EBV) reactivation. The diagnosis of leptospirosis was performed using four different diagnostic methods. Sera were analyzed with an in-house IgM and IgG enzyme-linked immunosorbent assay (ELISA) and indirect haemagglutination assay (IHA). Microscopic agglutination test (MAT) was done using 17 reference strains comprising 14 serogroups and 17 serovars. Polyvalent EBV-IgG analysis, EBV-IgG/IgM/IgA western blot analysis as well as quantitative EBV polymerase chain reaction (PCR) were performed. Leptospira IHA showed an initial titer of 1:640 (cut-off 1:320), leptospiral IgG was negative, but IgM was positive. MAT was negative at that time for all 17 strains analyzed. One week later, leptospirosis IHA titer increased to 1:20,480. Leptospiral IgG was now positive, -IgM remained positive and urine was tested negative for leptospiral DNA. The MAT showed positive results for L. interrogans serovar Bataviae, serovar Copenhageni, serovar Pyrogenes and L. borgpetersenii serovar Serjoe. During follow-up examinations, both the leptospiral IgM and IgG remained positive and MAT showed positive results for L. interrogans of different serovars. EBV IgA immunoblot taken at admission was positive for VCA-p18, quantitative EBV-PCR showed an EBV viral load of 2.8E3 copies/ml indicating acute EBV-reactivation. Leptospirosis represents a neglected and re-emerging disease which is difficult to diagnose since Leptospira-PCR from whole blood or urine is frequently negative in the case of early empiric antibiotic treatment. EBV-reactivation might represent a severe complication in Weil's disease which potentially aggravates clinical manifestations of leptospirosis including hepatitis, nephritis, and rhabdomyolysis. Thus, there might be a need for peripheral blood EBV-PCR and EBV blotting in patients suffering from complicated Weil syndrome, also in terms of the choice of antibiotic treatment.

Association between Epstein - Barr Virus (EBV) and Breast Cancer

Background: Breast cancer is the most common malignancy in females worldwide. Several etiological factors including environmental factors have been recognized for breast cancer. Epstein Barr virus as a viral etiological factor has been proposed. So far, several studies have investigated the relationship between development of breast cancer and Epstein Barr virus, but few have been done in Iran. The aim of this study was to determine whether there was an association between EBV infection and female breast cancer in Iran. Materials and Methods : We analyzed paraffin embedded breast tissue specimens by polymerase chain reaction (PCR) including breast cancer specimens (as case group) and breast fibroadenoma specimens (as control group). PCR was performed to amplify specific sequences of EBV. Results : From 130 cases of breast samples, 67 cases of breast cancer tissues and 41 cases of breast fibroadenoma tissues had adequate quality and quantity of DNA to detect EBV. PCR for EBV was positive in 4 invasive ductal carcinoma specimens (7.3%) and only one of the fibroadenoma specimens (2.4%). No significant association was found between EBV infection and invasive ductal carcinoma (p> 0.05). Also, patient’s age and histological grade of IDC were not correlated with EBV infection (p>0.05). Conclusion : We observed no etiologic association between EBV infection and invasive ductal carcinoma of female breast in our regions; however, further studies are required to elucidate this association.

Prevention of Posttransplant Lymphoproliferative Disorder in Pediatric Patients With a Liver Transplant.

This study sought to evaluate in pediatric liver transplant recipients the effects of hybrid antiviral therapy on the rate of posttransplant lymphoproliferative disorder. All pediatric patients (87 cases) who had undergone a liver transplant between April 2011 and March 2012 took part in the study and received hybrid antiviral treatment (case group). Epstein-Barr virus polymerase chain reaction was monitored intermittently. The results were compared to those of a historical control group including 117 pediatric patients who received a liver transplant between April 2009 and March 2011. Follow-up was 27 to 47 months in the control group and 12 to 26 months in the case group. Posttransplant lymphoproliferative disorder occurred in 12 patients in control group (10.2%) and 5 patients in case group (5.7%) (P = .249). Of 12 cases of posttransplant lymphoproliferative disorder, death occurred in 5 cases in the control group (41.7%), while no posttransplant lymphoproliferative disorder-associated death was seen in the case group (P = .086). Although hybrid antiviral treatment did not result in a statistically significant decrease in posttransplant lymphoproliferative disorder and posttransplant lymphoproliferative disorder-associated mortality rates, considering the limited number of posttransplant lymphoproliferative disorder cases in this study, this decrease may be interpreted as noticeable, and we advise using this strategy for pediatric patients undergoing a liver transplant.

Epstein-barr virus-negative post-transplant lymphoproliferative diseases: three distinct cases from a single center.

Three cases of Epstein-Barr virus (EBV)-negative post-transplant lymphoproliferative disease that occurred 6 to 8 years after renal transplantation are reported. The patients respectively had gastric mucosa-associated lymphoid tissue lymphoma, gastric diffuse large B-cell lymphoma, and atypical Burkitt lymphoma. Absence of EBV in the tissue samples was demonstrated by both in situ hybridization for EBV early RNA and polymerase chain reaction for EBV DNA. Patients were treated with reduction in immunosuppression and combined chemotherapy plus an anti-CD20 monoclonal antibody, rituximab. Despite the reduction in immunosuppression, patients had stable renal functions without loss of graft functions. The patient with atypical Burkitt lymphoma had an abnormal karyotype, did not respond to treatment completely, and died due to disease progression. The other patients are still alive and in remission 5 and 3 years after diagnosis, respectively. EBV-negative post-transplant lymphoproliferative diseases are usually late-onset and are reported to have poor prognosis. Thus, reduction in immunosuppression is usually not sufficient for treatment and more aggressive approaches like rituximab with combined chemotherapy are required.