Peripheral Blood EBV-DNA Research Articles

Investigation of lymphoma patients' EBV infection status

This study was purposed to investigate the EBV infection status of lymphoma patients from January 2008 to April 2012 in the First Hospital of Peking University. All the candidates have been detected for EBV which was either peripheral blood EBV DNA or ISH EBER in pathology from January 2008 to April 2012. The information on their sex, age, pathological type, peripheral blood EBV DNA and ISH EBER was collected, the positive rate of different EBV tests was studied, and the different characteristics of the EBV(+) and EBV(-) group were also explored. And Kaplan-Meier and Cox survival analysis was applied to investigate the EBV's effect on overall survival of these patients. The results showed that among 169 lymphoma patients, the positive rates of EBV EBER in extranodal NK/T cell lymphoma, angioimmunoblastic T cell lymphoma and peripheral T-cell lymphoma were 84.8%, 72.7% and 40.0%, respectively, and were ranged as the top three. The positive rate of EBV in DLBCL was relatively lower (16.7%) than that in above three types of lymphoma. The positive rate of peripheral blood EBV DNA of the elderly EBV(+) DLBCL was 50%. One out of 10 HL patients was subjected to EBER detection, the result of which was positive. The positive rate of peripheral blood EBV DNA of HL was 10%. Both the T cell lymphoma proportion and the rate of B symptom were higher in EBV(+) group than in EBV(-) group. In all the EBER(+) cases, the difference of OS between EBV(+) and EBV(-) patients was statistically significant. In multiple-factor survival analysis, peripheral blood EBV DNA positive was an independent risk factor for poor prognosis in the patients with lymphoma. It is concluded that EBV is closely related to extranodal NK/T cell lymphoma, angioimmunoblastic T cell lymphoma and peripheral T-cell lymphoma. Peripheral blood EBV DNA positive is an independent risk factor for poor prognosis in lymphoma patients.

EBV-DNA In Peripheral Blood Of Patients With Diffuse Large B Cell Lymphoma: Associations With Patient Characteristics and Outcome

Infection of EBV has been linked to the etiology of a number of lymphoproliferative diseases, and EBV+ diffuse large B cell lymphoma (DLBCL) of the elderly has been more recently described as a distinct entity. The viral load of EBV-DNA in plasma can be an indicator for EBV-association in some lymphoma types, as NK/T-cell lymphoma and Hodgkin lymphoma, and a marker for disease activity associated with negative prognosis. Here, we studied whether EBV-DNA in whole blood (WB), mononuclear cell fraction (MNC) and plasma of patients with DLBCL at diagnosis could be an indicator for EBV-association and a prognostic marker in patients treated with immunochemotherapy (R-CHOP).We analysed 247 patients with DLBCL (median age 64 years, range 15-92 years; 118 females, 129 males), diagnosed between 2006 and 2013. All patients were HIV-negative. EBV was detected using a commercial real-time PCR kit (BioQuant EBV, Biodiversity, Brescia, Italy) in peripheral blood (PB) compartments (WB n=240, plasma n=55, and MNC n=52). Lymph node samples from 61 DLBCL patients were analyzed for EBV infection through in situ hybridization for EBV-encoded small RNAs (EBER).EBV was detected in 58 of 240 WB samples (24%). The copy number varied between 2 x 102 and 4.9 x 106 copies/ml. The presence and copy number of EBV in WB and MNC were correlated (p<0.01, respectively), while there was no correlation to the detection of EBV in plasma. We did not find any association between the presence or viral load of EBV-DNA in any blood compartment and the presence of EBV in the lymphoma cells of 61 patients studied with EBER–ISH (11 patients EBER positive). Elderly patients (>60 years of age) had more often EBV in peripheral blood (30% vs 17%), and the viral load was higher in patients aged > 60 years (p<0.01). The presence of EBV-DNA in PB was also strongly associated with a positive serology for Hepatitis C virus (HCV): 20/221 patients (8.8%) were HCV+, and 11/20 patients (55%) harbored EBV-DNA in PB, while in the HCV- patients the frequency of EBV-positivity in PB was 21% (42/201) (p=0.002). As a positive serology for HCV is associated with age > 60 years (p=0.02), we performed a logistic regression analysis including age and HCV serology as parameters determining the EBV viral load in PB. In this analysis, HCV serology, but not age remained a significant factor for EBV viral load (p=0.004). Presence and viral load of EBV in PB was not related to other characteristics as gender, stage, performance status, LDH level, and IPI. In an univariate analysis including 201 patients treated with R-CHOP, the presence of EBV-DNA in WB was associated with a significantly shorter event-free survival (EFS): 67% (50%-79%, 95% C.I.) versus 80% at 2 years (p<0.04). Correcting for IPI and HCV serology in a multivariate Cox analysis, the presence of EBV-DNA in PB retained its prognostic significance (HR 1.9; 95% C.I., 1.07-3.43; p<0.03).Our findings suggest that EBV can be frequently detected in peripheral blood at DLBCL diagnosis, that is not a surrogate marker for EBV-status in DLBCL, but associates with a worse outcome. Further studies are needed to explore the mechanisms of expansion of EBV-positive cells in patients with DLBCL. Disclosures:No relevant conflicts of interest to declare.

Subcutaneous panniculitis-like T-cell lymphoma presented with full facial edema, hemophagocytic syndrome and Epstein-Barr Virus infection:A case report

BackgroundSubcutaneous panniculitis-like T-cell lymphoma (SPTCL) is a rare and distinct type of T-cell lymphoma. It is frequently associated with hemophagocytic syndrome (HPS), whose etiologic factors always involve Epstein-Barr Virus(EBV) infection. Although EBV is absent in many cases of sporadic SPTCL, we found a case of SPTCL with full facial edema, HPS and EBV infection as the first symptom. Patient and treatmentA 36-year-old female had presented with fever, full facial edema for 20 days before hospitalization. Remarkable abnormalities were found as hypersplenotrophy, pancytopenia, hypofibrinogenemia, high ferritin and soluble interleukin-2-receptor level, hemophagocytosis in bone marrow smear, which indicated HPS. Detection by PCR of EBV-DNA in peripheral blood was 9.73×104 copies. On the next day of her hospitalization, HLH-2004 protocol (VP-16+dexamethasone+cyclosporine A) was performed. Her temperature fell to normal on the 4th day. Full facial edema vanished gradually in the following days. After 8 weeks' treatment, all the abnormalities in the laboratory examination returned to normal. Despite of cyclosporine A and prednisone were used as continual treatment, facial edema and fever relapsed accompanied by multiple skin lesions on her back, upper and lower limbs 4 months later. Reexamination of the laboratory index related above were abnormal once again. Copy number of EBV-DNA was 5.52×105. Histologic and immunohistochemical features of tissue biopsy from the skin lesion on her right arm revealed the evidence of SPTCL. Then, she was treated by asparaginase plus DICE regimen. The facial edema and skin lesion vanished gradually during the course of treatment. Now (one month after the first course of chemotherapy), the patient is in her second course of asparaginase plus DICE regimen and SPTCL remains remission. ConclusionThis was not a typical case of SPTCL. The patient presented with overt skin lesion 4 months after the first symptom of full facial edema, which suggested SPTCL is a heterogenous disease with variable presentations. Awareness of this rare entity is critical to both ensure timely diagnosis and treatment. In addition, the pathogenesis of SPTCL remains unclear. Some malignant lymphomas are induced by chronic inflammation due to viral or bacterial infection. It is well known that EBV infection often causes malignant lymphomas of B-cell type, particularly in elderly peoples and immunocompromised individuals. Although EBV is absent in many cases of sporadic SPTCL, the virus appears to be implicated in the etiology of variant SPTCL occurring in the Asian population. Disclosures:No relevant conflicts of interest to declare.

L-Asparaginase Monotherapy for T- and NK-Cell Type of Chronic Active Epstein-Barr Virus Infection in Adults: A Pilot Study

Abstract 4953Chronic active Epstein-Barr virus infection (CAEBV) is a rare and fatal disorder. In Japan and East Asia, most CAEBV patients are the T- or NK-cell-infected type and they have been resistant to the current chemotherapies.L-asparaginase (L-asp) is a reagent that has a well-known effect on extranodal NK-/T-cell lymphoma nasal type (ENKL) even as a single reagent. CAEBV is one of the EBV-positive T- or NK-cell lymphoproliferative diseases (EBV-T/NK-LPD) as is ENKL. In addition, L-asp is not influenced by P-glycoprotein, which is expressed in EBV-infected T-or NK-cells of CAEBV (our unpublished data). In this pilot study, therefore, we investigated its effects on T- and NK-cell type of CAEBV.Adult patients of CAEBV without severe organ dysfunction were enrolled. CAEBV was diagnosed according to the following criteria: persistent infectious mononucleosis-like symptoms, elevation (>1 X102 copies/μg DNA) in peripheral blood EBV-DNA titer (pEBV-DNA), and the presence of EBV-infected T- or NK-cells. To detect infected cells, we isolated peripheral mononuclear cells and divided them into CD19-, CD4-, CD8-, or CD56-positive fractions using antibody-conjugated magnetic beads. EBV-DNA of each fraction was quantified using a real-time quantitative polymerase chain reaction (PCR) assay. The treatment protocol involved 7 administrations of 6000 U/m2 L-asp every other day. The response was defined by the decrease of pEBV-DNA 1 month after treatment completion. We also performed quantification of asparagine synthetase (AS) in EBV-infected cells of the patients.The results were summarized in the table. Between February 2010 and September 2010, 5 females were enrolled. Two patients were previously treated with the regimen comprising cyclosporine A, prednisolone, and VP16. One patient had EBV-positive cell infiltration of muscles with an elevation of LDH and CK. The mean titer of EBV-DNA in peripheral whole blood was 1.2 × 105 copies/μg DNA.Three patients completed the treatment. pEBV-DNA decreased in 1 patient with a reduction rate of 0.09 with improvement of clinical symptoms, but increased 2.7-fold in another patient and remained almost unchanged in the other. The response rate was 20% (1/5). Of the patients whose treatment was discontinued, one showed progression of the nasal lesion with pEBV increasing 1.5-fold, and the other had a dystonic attack on day 11, and the 2 remaining administrations were stopped although the muscle lesions were improved.Several adverse events (AE) were detected, including liver dysfunction (grade 2 and 3) in 2 patients and neutropenia (grade 3) in 1 patient. One patient had a dystonic attack as described earlier. The brain MRI showed no lesion in the central nervous system that could have caused this attack. Because the patient had been diagnosed with dystonia before she developed CAEBV, the attack was considered not to be directly attributable to L-asp.We examined AS mRNA levels in the EBV-infected cells. mRNA level was low in the patient who achieved pEBV-DNA decrease. As shown in the table, however, we could not find a significant relationship between the effect and AS levels.In general, the effect of L-asp on CAEBV was limited and the rate of AE was high. However, L-asp may have responses in some patients. CAEBV has a diverse phenotype of clinico-pathological findings and associated symptoms. Treatment for CAEBV needs to be planned and evaluated according to the subtype of the disease. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

The Viral Load of Epstein–Barr Virus (EBV) DNA in Peripheral Blood Predicts for Biological and Clinical Characteristics in Hodgkin Lymphoma

The Epstein-Barr virus (EBV) is present in the malignant Hodgkin/Reed-Sternberg (HRS) cells of 20% to 40% cases of Hodgkin lymphoma (HL) in Western countries. We were interested in the detection and quantification of cell-free plasma EBV-DNA as an indicator of biological and clinical characteristics in EBV-associated HL. EBV was detected in peripheral blood compartments (whole blood, plasma, and mononuclear cells) at diagnosis by real-time PCR for the EBNA (EB nuclear antigen) region (n = 93) and in HRS cells by in situ hybridization for EBV-encoded small RNAs (EBER; n = 63). These data were correlated to histological and clinical characteristics, EBV serology, circulating cell-free DNA, and interleukin (IL)-6 levels. Detection of EBV-DNA in plasma had a high specificity (90%), but a relatively low sensitivity (65%) to predict for EBV association. The viral load was higher in patients with advanced stage disease, older age in the presence of B-symptoms, and international prognostic score more than 2. The presence of EBV in HRS cells and higher plasma EBV-DNA copy numbers correlated to an increased frequency of tumor-infiltrating CD68+ macrophages in lymph node biopsies. Plasma EBV-DNA load correlated to circulating cell-free DNA and IL-6 levels, and inversely correlated to lymphocyte counts and EBNA1 antibody titers. Although the presence of EBV-DNA in peripheral blood cannot be regarded as a surrogate marker for EBER, the plasma EBV-DNA load at HL diagnosis is an indicator of disease activity and biological characteristics associated with negative prognosis. Moreover, the inverse correlation to EBNA1 antibody titers and lymphocyte counts may indicate a reduction in immunosurveillance, favoring the expansion of EBV-HRS cells in HL.

Multicenter comparative study of Epstein–Barr virus DNA quantification for virological monitoring in transplanted patients

Background EBV-related post-transplant lymphoproliferative diseases are usually accompanied by increased EBV DNA in peripheral blood. Monitoring EBV DNAemia is the basis for weighing decisions regarding initiation of pre-emptive or anti-EBV-related tumor therapy. However, the definition of clinically relevant cut-off values is hampered by the lack of standardization in EBV DNA testing. Objectives To estimate inter-laboratory variability and to evaluate the impact of different matrices in EBV DNA load determination in Italian laboratories involved in monitoring of virus infections in transplanted patients. Study design Two different proficiency panels were distributed among seven centers: the first contained cell-associated and cell-free EBVs; the second was prepared by spiking both cell-associated and cell-free EBVs in EBV DNA-negative whole blood from EBV seropositive healthy donors. Samples were extracted and amplified with different methods. Intra-laboratory and inter-laboratory variabilities was evaluated. Results 337 EBV DNA determinations were performed. Sensitivity was 100% for both panels, specificity was 100% for the first and 74% for the second panel, where whole blood was utilized as the matrix. Discrepant results in the second panel were restricted to samples containing low copy numbers. Quantification fell within ±0.5 log in 73% of the determinations. Values for cell-associated samples tended to be more heterogeneous than those obtained from cell-free samples. Good overall linearity was observed for each sample type; inter-laboratory variability ranged from 4.71% to 12.86%. Conclusions The results of this multicenter study indicate that EBV DNAemia may be reliably quantified by different laboratories using a variety of commercial and in-house molecular assays.

Quantitation of EBV-DNA In Peripheral Blood In Hodgkin Lymphoma: Associations with Other Biomarkers and Patient Characteristics

AbstractAbstract 2678The Epstein-Barr virus (EBV) is present in the malignant Hodgkin/Reed-Sternberg (HRS) cells of 20–40% cases of Hodgkin lymphoma (HL) in western countries. Detection and quantitation of cell-free plasma EBV-DNA has been proposed as a biomarker in EBV-associated malignancies, including HL. We analyzed the presence of EBV in HRS cells, peripheral blood compartments, and correlated these data to EBV serology, other circulating biomarkers and clinical characteristics. In a single institution cohort of 93 HL patients, EBV-DNA was quantitated using real-time PCR for the EBNA region in whole blood (WB, n=69), plasma (PL, n=75) and peripheral blood mononuclear cells (MNC, n=74). Cell-free DNA levels were determined using a real-time PCR for the globin gene (n=84), cytokine concentrations were measured in 85 patients. EBV status of HRS cells was studied using in-situ hybridization with EBER. EBV-DNA was positive in 32% WB, 21% PL and 20% MNC at HL diagnosis, with concordant WB and PL results in 86% of cases. EBER was present in HRS cells in 32% of cases. EBV-DNA copy number over 1500/ml was predictive for the presence of EBER in HRS cells in 86% and 88% WB and PL, respectively, while EBV copy numbers <1500 had only 55% and 57% PPV for WB and PL. Older patients (>50 years) were frequently EBER-positive (66%), with higher EBV copy numbers (median 12800/ml), while younger patients (<50 years) were less frequent EBER-positive (23%), and copy numbers were lower (median 1200/ml) (p=0.01). EBER-positive HL cases frequently had EBNA-1 antibody titers < 100 U/ml when compared to EBER-negative cases (44% vs. 13%, p=0.01), and there was a trend for a negative correlation between EBV copy number and EBNA1 titers (p=0.07). In patients with EBER+ HL the plasma EBV-DNA copy number correlated to the level of circulating cell-free DNA (rho=0.65), IL-10 levels (rho=0.87), IL-6 levels (rho=0.74) and inflammation parameters, including CRP (rho=0.79) and ESR (rho=0.57) (all p<0.02), and were higher in stage IV disease (p=0.02). In conclusion, a high EBV-copy number in whole blood and/or plasma at diagnosis is a good indicator for EBV-association of HL, and EBV copy numbers strongly correlate to other biological parameters of disease activity. Moreover, the inverse correlation between EBV copy number and EBNA1 antibody titers may indicate a reduction in immune surveillance favoring the expansion of EBV. Disclosures:No relevant conflicts of interest to declare.

Clinical significance of clonality and Epstein‐Barr virus infection in adult patients with hemophagocytic lymphohistiocytosis

We assessed the clinical significance of T or B cell clonality and Epstein-Barr virus (EBV) infection in adult patients with hemophagocytic lymphohistiocytosis (HLH) to identify factors related to prognosis. A total of 30 adult patients with diagnosed HLH were included in the study. In all patients, EBV-DNA in peripheral blood was examined by quantitative real-time polymerase chain reaction and bone marrow cells were examined for clonal rearrangement of T cell receptor gamma(TCRG) and immunoglobulin heavy chain (IGH) genes. TCRG clones were detected in 10 patients (33.3%) and IGH clones were detected in 8 patients (26.7%). We found no correlation between clonality and patient outcome. The patients less than 1,000 copies (mL)21 of EBVDNA showed a significantly higher clinical response (P 5 0.008) and longer overall survival (P 5 0.01) than those with high viral load of EBV-DNA. Our results suggest that TCRG and IGH rearrangement do not have any clinical significance in adult patients with HLH, but that high viral load of EBV-DNA may be a risk factor for poor outcomes. In HLH, high viral load of EBV-DNA should thus suggest a prompt approach with aggressive therapeutic interventions.

Prospective Measurement of EBV-DNA in Plasma and Peripheral Blood Mononuclear Cells of Extranodal NK/T-Cell Lymphoma, Nasal Type.

Abstract 135 Background:Peripheral blood of patients with extranodal NK/T-cell lymphoma, nasal type (ENKL) contains fragmented Epstein-Barr virus (EBV) DNA. Measurement of the circulating viral DNA load has been reported to be useful for the diagnosis, monitoring and prognostication of the disease. However, there are two different subjects for analysis, plasma/serum component and mononuclear cells (MNC). Most reports do not compare both samples from the same patients, and are biased by a retrospective manner of accrual. Therefore, it remains unclear which samples are more useful subjects. Materials and Methods:To evaluate clinical significance of peripheral blood EBV-DNA copy number for ENKL, we conducted a prospective study to analyze EBV-DNA with quantitative polymerase chain reaction. Inclusion criteria are as follows: (1) Diagnosis of ENKL or aggressive NK-cell leukemia by biopsy or cytology. (2) Patients without other serious complications and those tolerable for chemotherapy and/or radiotherapy. (3) No prior history of chemotherapy or radiotherapy. (4) Patients with written informed consent. Primary endpoint was a prognostic value of EBV-DNA copy number to predict 2-year overall survival. Secondary endpoints were comparison of EBV-DNA copy number and pretreatment characteristics and prognostic capability of EBV-DNA during/after treatments. Three times of analysis, pre-treatment, mid-treatment, post-treatment, at central laboratory was to be performed for each patient. Results:A total of 33 patients were registered from June 2004 to March 2007. All patients were diagnosed with ENKL and male/female ratio was 21/12. The median age was 56 years old, ranging from 18 to 81. 19 patients were in stage IE, 3 in stage IIE, and 11 in stage IV. 13 patients had B-symptom and 14 had elevated serum LDH level. ECOG performance status was low (0–2) in 29 patients, but was 3 in 2 patients. IPI was high-intermediate/high in 11. First line treatment included concurrent chemoradiotherapy in 16, radiotherapy followed by chemotherapy in 8, and chemotherapy alone in 6. Pretreatment MNC and plasma EBV-DNA were detectable in 6 and 14 patients, respectively. The maximum copy numbers were 780 copies/microgram DNA for MNC and 71000 copies/mL for plasma. Significant correlation was observed between mononuclear and plasma EBV-DNA copies (r = 0.8741, P < 0.0001). Plasma EBV-DNA well correlated with pretreatment clinical stage (P = 0.02), presence of B-symptom (P = 0.02), ECOG PS (P = 0.02), serum LDH level (P = 0.05), and soluble IL-2 receptor (P < 0.0001), but not with regional node involvement (P = 0.17), nasal vs. nasal-type (P = 0.16), and serum C-reactive protein (P = 0.29). Among 28 patients evaluable for response, 21 patients responded (CR/PR) to the first line treatment. Mean plasma EBV-DNA copy number before treatment was significantly higher in non-responders compared to responders (16472 copies/mL vs 2645 copies/mL, P = 0.02). 2-year overall survival was 69.7%. The median follow-up of patients was 2.9 years. Clinical stage, performance status, pretreatment plasma EBV-DNA and pretreatment mononuclear cell EBV-DNA were significant prognostic factors for overall survival by univariate analysis. Multivariate analysis showed clinical stage (hazard ratio = 9.0, 95% confidence interval: 1.8–45.0) and pretreatment plasma EBV-DNA (hazard ratio = 10.6, 95% confidence interval: 1.3–87.0) were significant prognostic factors. 2-year overall survival of plasma EBV-DNA positive and negative patients was 42.9% and 94.4%, respectively (Figure, P = 0.0009). [Display omitted] Conclusions:Our study shows pretreatment plasma EBV-DNA copy number is a good indicator for both response to treatment and overall survival. Measurement of the plasma EBV-DNA is useful for prospective clinical trials and general practice. Disclosures:Oshimi:Eisai Pharmaceutical Company: Employment.

Quantificazione mediante PCR dell’EBV-DNA da biopsie cutanee di pazienti con linfomi cutanei primitivi (micosi fungoide e sindrome di Sèzary)

Mycosis fungoides (MF), the most indolent form of CTCL, originates from a clonal expansion of epidermotropic helper/memory T cells. Sezary syndrome (SS) is a rare primay epidermotropic cutaneous T-cell lymphoma in leukemic. The aetiopathogenesis of MF and SS remains obscure despite several investigations. Infectious, environmental and genetic factors have been implicated as potential aetiological agents. The studies investigating the role of EBV in CTCL present conflicting results. The different sensitivities of the technical methods used in the evaluation of the presence of viral DNA or virus-related antigens make comparison of the results difficult. The aim of this study was to retrospectively evaluate the EBV-DNA load in skin biopsies from MF and SS patients by a highly sensitive (1-10 EBV-DNA copies/reaction) quantitative-competitive PCR (QC-PCR) developed in our lab to better asses the relationship between EBV and CTCL. Skin biopsies were obtained from 21 MF and 10 SS patients; skin biopsies from a 8 patients with inflammatory skin disease were used as controls. EBV-DNA was detected in 70% of biopsies from SS patients vs. 0% of MF patients. No control patients resulted EBV-DNA positive, as expected. In addition, in SS patients, the survival from diagnosis is lesser in EBV-positive patients vs.EBV-negative patients even if not statistically significant.We are going to investigate the presence of EBV-DNA in peripheral blood of a larger number of patients and to evaluate the pattern of viral genes expression, to better assess the aetiopathogenetical role of EB virus in this kind of neoplasies.

Epstein–Barr virus DNA load and seroconversion in pediatric renal transplantation with tacrolimus immunosuppression

EBV infection is one of major complications arising in pediatric patients who have undergone renal transplantation. A strong correlation between the grade of immunosuppression and the development of PTLD, one of the most severe EBV-associated diseases, has been recognized. In this study, we monitored the serologic profile in conjunction with peripheral blood EBV-DNA load of 32 children who underwent renal transplantation with tacrolimus as an immunosuppressant. Six patients were EBV-seronegative (EBV-) before the transplantation, and the mean DNA load in the EBV- group was significantly higher than that in the EBV-seropositive (EBV+) group. Seroconversion occurred in five of these patients in a mean period of 22 weeks after the transplantation. The EBV-DNA load in the EBV+ group was maintained at a low level for a year, whereas it increased rapidly to over 1 x 10(5) copies/mL in two patients in the EBV- group three to seven months after the transplantation, which corresponds to the timing of seroconversion, and one of them developed PTLD. These observations suggest that the close monitoring of the EBV-DNA load, along with longitudinal observation of seroconversion, is essential in pediatric renal transplantation, particularly for younger children who are more likely to be EVB-.

Analysis of Epstein-Barr virus reservoirs in paired blood and breast cancer primary biopsy specimens by real time PCR.

IntroductionEpstein-Barr virus (EBV) is present in over 90% of the world's population. This infection is considered benign, even though in limited cases EBV is associated with infectious and neoplastic conditions. Over the past decade, the EBV association with breast cancer has been constantly debated. Adding to this clinical and biological uncertainty, different techniques gave contradictory results for the presence of EBV in breast carcinoma specimens. In this study, minor groove binding (MGB)-TaqMan real time PCR was used to detect the presence of EBV DNA in both peripheral blood and tumor samples of selected patients.MethodsPeripheral blood and breast carcinoma specimens from 24 patients were collected. DNA was extracted and then amplified by MGB-TaqMan real time PCR.ResultsOf 24 breast tumor specimens, 11 (46%) were positive for EBV DNA. Of these 11 breast tumor specimens, 7 (64%) were also positive for EBV DNA in the peripheral blood, while 4 (36%) were positive for EBV DNA in the tumor, but negative in the blood.ConclusionEBV was found at extremely low levels, with a mean of 0.00004 EBV genomes per cell (range 0.00014 to 0.00001 EBV genomes per cell). Furthermore, our finding of the presence of EBV in the tumor specimens coupled to the absence of detection of EBV genomic DNA in the peripheral blood is consistent with the epithelial nature of the virus. Because of the low levels of viral DNA in tumor tissue, further studies are needed to assess the biological input of EBV in breast cancer.

496. PCR 法を用いた EB ウイルスの DNA 検出 : 末梢血からの DNA 抽出について(核医学 試料測定-2)

496. PCR 法を用いた EB ウイルスの DNA 検出 : 末梢血からの DNA 抽出について(核医学 試料測定-2)