Circulating Epstein-Barr Virus DNA Research Articles

The Dynamic Change in Plasma Epstein-Barr Virus DNA Load over a Long-Term Follow-Up Period Predicts Prognosis in Nasopharyngeal Carcinoma.

The current study was designed to investigate the changes in the circulating Epstein-Barr virus DNA load (EBV DNA) at various time points before and after treatment and its clinical significance in nasopharyngeal carcinoma (NPC). A total of 142 patients with NPC were prospectively enrolled in this study. The plasma EBV DNA concentration was measured before and after treatment using qPCR. The prognostic values of the EBV DNA load were analyzed using the Kaplan-Meier and Cox regression tests. Following multivariate analysis, our data showed that high pre-EBV DNA loads were associated with significantly poorer distant metastasis free survival (DMFS) and progression free survival (PFS); detectable end-EBV DNA loads were associated with significantly worse loco-regional recurrence free survival (LRRFS) and PFS, and the detecTable 6 months-post-EBV DNA loads were associated with significantly poorer overall survival (OS), DMFS and PFS (p < 0.05). Additionally, combining the pre-EBV DNA load and the stage of the disease, our results showed that patients at stage III-IVA with a low pre-EBV DNA load had similar survival rates as patients at stage II with a low or high pre-EBV DNA load, but had better survival rates than those at stage III-IVA with a high pre-EBV DNA load. Taken together, we showed that the change of the EBV DNA load measured at several time points was more valuable than at any single time point for predicting patients' survival for NPC. Furthermore, combining the pre-EBV DNA load and the TNM classification could help to formulate an improved prognostic model for this cancer.

Prognostic significance of circulating Epstein-Barr virus DNA in pulmonary lymphoepithelioma-like carcinoma: A meta-analysis and validation study.

In small-scale studies, circulating Epstein-Barr virus (EBV) DNA levels have prognostic value in patients with pulmonary lymphoepithelioma-like carcinoma (LELC). Therefore, we performed a comprehensive meta-analysis to evaluate the prognostic significance of circulating EBV DNA levels in patients with pulmonary LELC. Studies that discussed the prognostic significance of circulating EBV DNA detection in pulmonary LELC were eligible for inclusion in this study. The overall survival (OS) and progression-free survival (PFS) were the primary outcomes. Pooled hazard ratio (HR), 95% confidence intervals (CIs), and pvalue were calculated to estimate the prognostic significance of EBV DNA levels. Additionally, we conducted a further observation using an independent cohort. The pooled HR and 95% CI of pretreatment EBV DNA levels for OS and PFS were 3.63 (95% CI: 2.90-4.55) and 2.88 (95% CI: 1.90-4.38), respectively. The pooled HR and 95% CI for Posttreatment EBV DNA levels for OS and PFS were 3.77 (95% CI: 2.96-4.80) and 3.52 (95% CI: 1.91-6.51, p < 0.001), respectively. The independent cohort showed similar results that patients with high pretreatment EBV DNA or positive posttreatment EBV DNA had significantly inferior PFS. Circulating EBV DNA levels provide prognostic values of survival and treatment response in pulmonary LELC patients.

Circulating Epstein-Barr virus DNA associated with hepatic impairment and its diagnostic and prognostic role in patients with gastric cancer.

Epstein–Barr virus (EBV) infection may affect all tissues and organs of the body. Little is known about the impact of this entity on its systematic incorporation in patients with gastric cancer (GC). This study enrolled a total of 113 GC patients with EBV infection (EBVaGC) and 167 GC patients without EBV infection (EBVnGC). It was found that the CRP levels (indicative of inflammatory status) were significantly increased in EBVaGC compared with those in EBVnGC (12.11 mg/L vs. 5.72 mg/L, P = 0.008), but WBC and neutrophils counts were similar in both groups (P > 0.05). Consistent elevations in the levels of liver enzymes, ALP and GGT, with incompatible alterations in ALT or AST were observed in EBVaGC. Slightly prolonged coagulation indices, PT and INR, and decreased albumin consistently suggested impaired synthesis capability of the liver in EBVaGC (all P < 0.05). The level of circulating EBV DNA was positively correlated with the level of GGT, tumor marker CA72-4 and the lymphocyte infiltration in tumor tissues (all P < 0.05). Of note, the EBV associated high-lymphocyte infiltrated tissues presented rich CD8 + T cells. Circulating EBV DNA further showed a predictive role in distinguishing EBVaGC from EBVnGC (AUC 0.79, 95% CI 0.73 to 0.85, P < 0.001), and was associated closely with better overall survival (HR 0.45, 95% CI 0.21 to 0.96, P = 0.039). EBV infection in patients with gastric cancer may be linked to hepatic impairment and immune response. Circulating cell-free EBV DNA is not only a biomarker for the screening of an EBV-related GC subtype but is also an independently prognosis factor for the long-term survival benefit in GC patients.

Relative Expression of mRNA BARF1 Epstein-Barr Virus from Tumor Tissue Biopsy in Formalin-Fixed Paraffin-Embedded in Nasopharyngeal Carcinoma

Nasopharyngeal carcinoma (NPC) is a malignant tumor originating from nasopharyngeal epithelial cells. NPC is endemic in some regions, especially in Southeast Asia countries. NPC is also a multifactorial disease involving environmental factors, genetic factors, and infection from Epstein-Barr virus (EBV). According to WHO classification, Undifferentiated NPC is histopathologically associated with EBV infection and categorized into WHO subtype 3. EBV has two phases in its infection cycle: the lytic and latent phases. The BARF1 gene is a mediator of the transition from the latent phase to the lytic phase. Previous studies suggest measurement of EBV mRNA activity at the primary tumor site in the nasopharyngeal reflects the pathogenesis of NPC rather than measuring circulating EBV DNA or serological diagnosis. This study aimed to determine the relative expression potential of BARF1 mRNA at different tumor stages in NPC patients as a predictor of NPC pathogenesis. This research design was a descriptive research method in the form of a cross-sectional study. The samples used were 22 patients diagnosed as NPC WHO class III at the Anatomical Pathology Section of Prof. Dr. Margono Soekarjo, Purwokerto, who met the inclusion criteria. The relative expression of BARF1 mRNA was carried out using the one-step real-time RT-PCR technique and then calculated using a formula of 2-ΔCt. The T-test was used to compare the relative expression of early and late-stage BARF1 mRNA. The relative expression of BARF1 mRNA in the late-stage advanced stage (n = 6; 0.708292-0.840177; med = 0.7164655) was increased compared to the early stage (n = 2; 0.708841-0.712423; med = 0. 710632).

Utilization of the lymph node-to-primary tumor ratio of PET standardized uptake value and circulating Epstein–Barr virus DNA to predict distant metastasis in nasopharyngeal carcinoma

Background and purposeTo determine the clinical impact of integrating Epstein–Barr virus (EBV) DNA and lymph node-to-primary tumor ratio (NTR) of positron emission tomography (PET) standardized uptake value (SUV) in predicting distant metastasis, such as distant metastasis-free survival (DMFS), in patients with nasopharyngeal carcinoma (NPC). Materials and methodsWe retrospectively reviewed patients diagnosed with non-disseminated NPC between 2010 and 2017. The optimal cut-off values of EBV DNA and SUV NTR were determined using receiver operating characteristic analysis. The prognostic values of SUV NTR and EBV DNA on DMFS and overall survival were evaluated using the Kaplan–Meier method. Univariate and multivariable analyses were performed using the Wald Chi-squared test and Cox proportional hazards regression, respectively. ResultsA total of 488 patients were included in the analysis. The median follow-up period was 61.6 months. The optimal cut-off values of EBV DNA and SUV NTR were 3377.5 copies per mL and 0.64, respectively. The five-year DMFS for patients with high vs low EBV DNA and SUV NTR levels were 64.9% vs 86.6% (p < 0.001) and 78.7% vs 87.4% (p = 0.021), respectively. In subgroup analysis, the high-risk group with high levels of pretreatment EBV DNA and SUV NTR had worse DMFS in either American Joint Committee on Cancer (AJCC) stage I–III or IVA–B (p = 0.001 and <0.001, respectively). Univariate and multivariable analyses showed the statistical significance of EBV DNA, SUV NTR, and their composite in DMFS (p < 0.001 for EBV DNA; p = 0.022 for SUV NTR; p < 0.001 for their composite). ConclusionThis study showed that EBV DNA and SUV NTR have independent and additive values as prognosticators for distant metastasis in patients with NPC, suggesting that these two individual factors, except the AJCC staging system, should be included in future studies.

Clinical features in salivary gland lymphoepithelial carcinoma in 10 patients: Case series and literature review.

ObjectiveLymphoepithelial carcinoma (LEC) accounts for 0.4% of malignant tumors of the salivary gland and 0.8% of parotid gland malignancies. Over the past 50 years, less than 300 cases have been reported in the literature. The purpose of this study was to investigate the characteristics of salivary gland LEC.MethodsWe retrospectively reviewed the medical records and analyzed clinical data obtained from 10 patients seen at our hospital between 2005 and 2020 with salivary gland LEC.ResultsAll patients presented with a self‐palpable, non‐tender, hard swelling, or lump near the jaw or infra‐auricular region. Most cases (n = 8) were of solitary tumors, and enhancing patterns on computed tomography mainly were homogenous (n = 8). Interestingly, eight patients tested positive for the Epstein–Barr encoding region in in situ hybridization. Still, only three patients had detectable circulating Epstein–Barr virus (EBV) DNA, and one patient had detectable EBV IgA. All patients underwent complete tumor resection, followed by radiotherapy, and six also underwent chemotherapy. Nine patients became disease‐free within 5 years, and one died due to disease 4 years after surgery.ConclusionAlthough rare and considered to be a high‐risk malignancy, LECs have favorable treatment outcomes. Circulating EBV DNA is still not considered a marker for preoperative assessment or postoperative treatment response. The role of EBV DNA requires further investigation.Level of Evidence: 4

Epstein-Barr Virus is Associated with Gastric Cancer Precursor: Atrophic Gastritis.

Background: About 10% of gastric cancer (GC) has been described to be Epstein-Barr virus (EBV) positive. Previous researches have described the association between EBV and GC. However, the association of EBV with atrophic gastritis (AG) is underrecognized. Our study aimed to investigate the relationship between EBV and AG and assess the influence of EBV on gastric function.Methods: A total of 468 pathologically-confirmed chronic gastritis patients underwent circulating EBV DNA test, include 271 non-atrophic gastritis (NAG) and 197 AG patients.Results: In this study, H. pylori infection rate was 33.3%, EBV infection rate was 40%, and co-infection rate was 15%. The EBV DNA-positive was significantly associated with AG (P=0.031, OR= 1.509, 95% CI 1.037-2.194), especially in H. pylori-negative subjects (P=0.044, OR=1.619, 95% CI 1.012-2.589). EBV DNA-positive patients had a lower pepsinogen I (PG I) / pepsinogen II (PG II) ratio (PGR) than EBV DNA-negative patients (P=0.0026), especially in the AG subgroup (P=0.0062). There was no significant association between EBV and H. pylori co-infection with increased risk of AG (P>0.05).Conclusion: EBV infection significantly increased the risk of AG, especially in H. pylori-negative patients. The circulating EBV DNA had a potential in predicting the risk of atrophic gastritis.

Cell-free EBV DNA as a biomarker during clinical management of nasopharyngeal carcinoma in a nonendemic region.

Nasopharyngeal carcinoma (NPC) is the most common malignant tumor of the nasopharynx. Although NPC is not endemic in India,higher incidences were observed in its North-Eastern regions particularly Sikkim, Nagaland, Manipur, and Mizoram. Early detection of NPC is difficult because the nasopharynx is not readily amenable to clinical examination and symptoms of NPC are nonspecific.The development of suitable biomarkers for early diagnosis of NPC as well as accurate monitoring of treatment response is needed urgently. In this exploratory pilot study, we have investigated the clinical significance of assessing plasma Epstein-Barr virus (EBV) DNA load at diagnosis and during treatment. We found that EBV DNA is detectable at diagnosis in the majority of patients with nonendemic NPC and the absolute copy number of circulating EBV DNA per milliliter increases progressively with the stage of the disease. The viral load declined significantly with induction chemotherapy and definitive chemoradiation but showed a sharp rise at relapse. Patients with EBV DNA levels ≥1500 copies/ml had a higher risk of disease progression or relapse when compared with patients who had EBV DNA <1500 copies/ml at baseline. Estimation of plasma EBV DNA may serve as an excellent noninvasive tool to monitor disease extent, response to therapy, and for better prediction of future relapse or progression-free survival in a nonendemic NPC patient population.

Markers of Epstein–Barr virus in clinical assessment of Russian patients with nasopharyngeal cancer

Introduction. Epstein–Barr virus (EBV) is equally widespread in the endemic and non-endemic world regions for nasopharyngeal cancer (NPC). High incidence of NPC in endemic countries and low in non-endemic countries suggest there are different mechanisms and conditions for tumor occurrence and, possibly, different clinical significance of EBV-associated markers. However, significance of these markers for determining NPC in non-endemic regions is still poorly understood. Objective – to determine clinical significance of titers of IgG/IgA antibodies to EBV capsid antigen and concentrations of the viral DNA in patients’ blood plasma as diagnostic and monitoring markers for NPC in a non-endemic region of Russia. Materials and methods. Titers of EB-specific antibodies were determined by indirect immunofluorescence, and concentration of the viral DNA in plasma was measured using a quantitative polymerase chain reaction in real time. Study group included patients with NPC (n = 96), and control group – blood donors (n = 171) and patients with other head and neck tumors (n = 33).Results. Titers of IgG/IgA antibodies to EBV capsid antigen, being an important diagnostic marker of nasopharyngeal cancer, did not always correlate with patients’ clinical condition. Humoral response to emerging events often delayed due to inertia of the immune system. Concentration of EBV DNA in patients’ blood plasma clearly reflected the dynamics of the pathological process: it decreased to background values in remission and increased while the disease progressed. In contrast to endemic regions, we did not find any correlation between the studied EBV markers and clinical manifestations of the disease, evaluated in accordance with the TNM classification (Tumor, Nodus and Metastasis).Conclusion. In non-endemic countries, such as Russia, serological and molecular markers of EBV can be successfully used for the primary diagnosis of NPC. However, for the disease monitoring, it is preferable to use the value of the concentrations of circulating EBV DNA, which, in contrast to the values of IgG/IgA antibody titers to VCA EBV, more accurately reflect the patient’s clinical condition.

Abstract IA01: Plasma DNA-based molecular diagnostics: Fragments, circles and beyond

Abstract My group has a longstanding interest in the diagnostic applications of cell-free DNA in plasma. For example, we have pioneered noninvasive prenatal testing (NIPT) and the use of plasma DNA for cancer screening. We believe that understanding the fundamental biology of circulating DNA is essential to further advance the field. In this regard, we have been particularly interested in the fragmentation patterns of cell-free DNA in plasma. We have shown that cell-free fetal DNA molecules in the plasma of pregnant women are shorter than those of maternal origin. The differential size profiles between circulating fetal and maternal DNA have enabled the use of plasma DNA size as an independent metric for NIPT. In the area of cancer liquid biopsy, we have used circulating Epstein-Barr virus (EBV) DNA as an example of tumor-derived DNA in plasma. Hence, circulating EBV DNA molecules are also short fragments of DNA. In another example, we have used hepatocellular carcinoma as a model system and have found that circulating tumor-derived DNA molecules have a shorter size profile than those of nontumoral origin. We are exploring enzymes that might be involved in the fragmentation of plasma DNA. We have shown that mice in which both copies of the Dnase1l3 gene have been knocked out have a larger size profile of circulating DNA. We have also developed a plasma DNA end motif metric to measure the effect of disruption of this nuclease system. In addition to the fragmentation of linear DNA in plasma, my group is also interested in exploring the presence of nonlinear DNA in plasma. Hence, we have used mitochondrial DNA (mtDNA) as a model system as mtDNA exists as a circular molecule inside mitochondria. Interestingly, we have observed the existence of both linear and circular mtDNA in plasma. Using liver and bone marrow transplantation models, we have shown that the circular form is predominantly of hematopoietic origin while the linear form is predominantly from the liver. Plasma mtDNA topologics thus holds information on the origin of circulating DNA. These studies therefore demonstrate some of the exciting findings that can be revealed through the study of plasma DNA fragmentomics and topologics. We believe that such new biologic understandings will drive new diagnostic applications. Citation Format: Yuk Ming Dennis Lo. Plasma DNA-based molecular diagnostics: Fragments, circles and beyond [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA01.

Epstein-Barr Virus DNA in Nasopharyngeal Carcinoma: A Brief Review.

Epstein-Barr virus (EBV) is a B-lymphocytes herpes virus and can transform B lymphocytes to malignant tumor cells if infected. Nasopharyngeal carcinoma (NPC) is strongly associated with EBV. Circulating EBV DNA in plasma has been recognized as an important biomarker of NPC. Much work has been done to validate the ability of circulating EBV DNA for screening, diagnosis, risk stratification, monitoring, and predicting prognosis. This chapter reviews the clinical progress of circulating cell-free EBV DNA in NPC.

High viral loads of circulating Epstein-Barr virus DNA copy number in peripheral blood is associated with inferior prognosis in patients with mantle cell lymphoma.

Mantle cell lymphoma (MCL) is a distinct subtype of B cell non-Hodgkin lymphoma. No research has yet documented to investigate the prognostic implications of Epstein-Barr virus (EBV) infection in MCL. The objective of this study was to determine whether EBV DNA load may influence the heterogeneity in the course of the disease in MCL patients. Eighty-eight MCL patients were retrospectively enrolled in the study. EBV DNA load was detected by real-time quantitative PCR for quantification. The univariate and multivariate Cox proportional hazards models were established for the estimation of prognostic factors. Twenty-seven patients were detected positive for EBV DNA and the median virus titer was 1.72×104 copies/mL (range, 8.20×102 to 4.14×105 copies/mL). With a median follow-up of 39 months (range, 9 to 120 months), patients in EBV DNA-positive group displayed unfavorable progression-free survival (PFS) (P=0.012) and overall survival (OS) (P=0.004) than patients in EBV DNA-negative group. Multivariate Cox regression analysis revealed that EBV DNA-positivity was an independent risk factor for both PFS (HR, 2.04; 95% CI, 1.07 to 3.92; P=0.031) and OS (HR, 2.68; 95% CI, 1.20 to 6.00; P=0.016). Reduction in EBV copies was significantly associated with therapy-response.Circulating EBV DNA load in whole blood proved to be a significant predictor of prognosis in patients with MCL, which needs further validation in large-scale clinical studies.

Methylation analysis of plasma DNA informs etiologies of Epstein-Barr virus-associated diseases

Epstein-Barr virus (EBV) is associated with a number of diseases, including malignancies. Currently, it is not known whether patients with different EBV-associated diseases have different methylation profiles of circulating EBV DNA. Through whole-genome methylation analysis of plasma samples from patients with nasopharyngeal carcinoma (NPC), EBV-associated lymphoma and infectious mononucleosis, we demonstrate that EBV DNA methylation profiles exhibit a disease-associated pattern. This observation implies a significant potential for the development of methylation analysis of plasma EBV DNA for NPC diagnostics. We further analyse the plasma EBV DNA methylome of NPC and non-NPC subjects from a prospective screening cohort. Plasma EBV DNA fragments demonstrate differential methylation patterns between NPC and non-NPC subjects. Combining such differential methylation patterns with the fractional concentration (count) and size of plasma EBV DNA, population screening of NPC is performed with an improved positive predictive value of 35.1%, compared to a count- and size-based only protocol.

Longitudinal circulating Epstein–Barr virus DNA response to induction chemotherapy and chemo-radiotherapy to identify biological phenotypes in EBV-associated nasopharynx of head and neck cancer.

6021 Background: Liquid biopsies have the utility for detecting minimal residual disease in several cancers, but its clinical utility for real-time treatment adaptation remains limited. We adopted Epstein-barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) as a model to investigate its potential. We characterize longitudinal response of circulating EBV DNA to induction chemotherapy (IC) and concurrent chemoradiotherapy (CCRT) in locally advanced NPC (LA-NPC), and investigate the association of complete biological response (cBR, undetectable cfEBV DNA) during treatment to prognoses. Methods: The medical records of 673 LA-NPC cases with serial EBV DNA measurements (pre-treatment, after each IC cycle, post-CCRT) were extracted. Cox regression and landmark analysis were used for survival analyses. Results: Four distinct phenotypes were identified based on their longitudinal cfEBV DNA response: 1) Early responders (200/673[29.7%]) achieved cBR post-IC1; 2) Intermediate responders (332/673[49.3%]) included patients with cBR post-IC2-4, and cBR post-CCRT after two IC cycles or following a temporary bounce (detectable reading following initial cBR); 3) Late responders (75/673[11.2%]) achieved cBR only post-CCRT after 3-4 IC cycles; 4) Treatment-resistant (66/673[9.8%]) patients demonstrated non-cBR post-IC+CCRT. These phenotypes were significantly correlated with prognoses, adjusted for pre-treatment EBV DNA load, N-category and chemotherapy intensity (AHRDFS = 3.46[2.01–6.25], intermediate; 7.50[4.24–14.77], late; 17.33[10.06–33.38], treatment-resistant vs. early responders, Pall &lt; 0.01). Interestingly, intermediate and late responders without cBR post-IC2 had inferior survival despite more IC (HRDFS = 1.83[1.17–2.84], &gt; 2 vs. &gt;2 IC cycles, P &lt; 0.01). For treatment-resistant patients, adjuvant chemotherapy following IC+CCRT reduced risk of distant metastasis (HRDMFS = 0.42[0.24–0.74], P &lt; 0.01). Conclusions: We propose investigate risk-adapted chemotherapy de-intensification and intensification strategies based on the four novel phenotypes, which could shape the individualized treatment of LA-NPC. Our study highlights the feasibility of liquid biopsy for real-time therapeutic adaptation.

Decreased oral Epstein-Barr virus DNA loads in patients with nasopharyngeal carcinoma in Southern China: A case-control and a family-based study.

The link of nasopharyngeal carcinoma (NPC) with Epstein‐Barr virus (EBV) has been established for decades. Although an abnormal high level of EBV sero‐antibody spectrum and cell‐free circulating EBV DNA loads were exhibited in NPC patients, oral EBV DNA loads, which are primarily responsible for the EBV transmission, has not been previously studied in NPC patients. We conducted an epidemiological study to measure the oral EBV loads, viral components, and the relationship with the serum antibody titers in a large case‐control population (968 cases and 1656 controls) and a family‐based population (91 cases and 165 unaffected family members). EBV DNA loads were detected by quantitative PCR approach targeting the BamHI‐W region. Although a large individualized variation existed, we still observed a decreased oral EBV DNA loads in the population of NPC patients compared to that of healthy controls (ORs were 1.00, 0.69, 0.62, 0.33 classified by the quartiles of viral loads, P trend < .001) and family members. In contrast, the elevated levels of oral EBV loads were present in asymptomatic males and elders, suggesting a different important source for EBV transmission. Notably, oral EBV loads were inversely associated with serum antibody titers of VCA‐IgA, EA‐IgA (All P trend < .001) in the cases but not in the controls. Our study provides the first epidemiological data of oral EBV loads and viral components in NPC patients and controls in the highest risk area of Southern China, indicating that NPC status is unlikely to be an important determinant of EBV transmission.

The Prognostic Role of Circulating Epstein-Barr Virus DNA Copy Number in Angioimmunoblastic T-Cell Lymphoma Treated with Dose-Adjusted EPOCH

PurposeDetermine the frequency and prognostic value of circulating Epstein-Barr virus (EBV) DNA copy number in angioimmunoblastic T-cell lymphoma (AITL) patients who were treated with dose-adjusted etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin (DA-EPOCH) regimens.Materials and MethodsSixty newly-diagnosed AITL patients were retrospectively enrolled in the present study. All patients were treated with DA-EPOCH regimen.ResultsTwenty-two subjects (36.7%) had a EBV DNA-positive test at diagnosis. EBV DNA‒positive patients were associated with lower lymphocyte-monocyte ratio (p=0.024). Median follow-up was 40 months (range, 14 to 100 months). The overall response rate for all the 60 AITL patents were 71.7% (95% confidence interval [CI], 58.6 to 82.5) with 3-year progressive-free survival (PFS) rate of 30.9%±6.1% and overall survival (OS) rate of 60.1%±6.6%. Not only did PFS estimation differ between the EBV DNA‒positive and EBV DNA‒negative group (hazard ratio [HR], 2.24; 95% CI, 1.15 to 4.35; p=0.006), but also worse OS was observed in the pretreatment EBV DNA‒positive group than in the EBV DNA‒negative group (HR, 2.74; 95% CI, 1.22 to 6.19; p=0.006). EBV DNA test positivity was independent prognostic marker for both PFS (HR, 2.17; 95% CI, 1.17 to 4.00; p=0.014) and OS (HR, 3.24; 95% CI, 1.48 to 7.11; p=0.004) after adjusting International Prognostic Index and prognostic index for AITL score. Reduction in EBV copies was significantly associated with therapy-response.ConclusionCirculating EBV DNA level was an important prognostic and monitoring marker for AITL patients who treated with DA-EPOCH regimens which cannot improve outcomes for AITL patients.

Analysis of an electronic methods for nasopharyngeal carcinoma: Prevalence, diagnosis, challenges and technologies

ContextNasopharyngeal Carcinoma (NPC) is a category of cancer in the head and neck regions, and this cancer prevails in the throat area amongst the pharynx and nasal cavity. This NPC represent major health problem in Malaysia, being the fourth most common tumor among Malaysians and third most common cancer disease among Malaysian men. NPC is emphatically connected with Epstein-Barr Virus (EBV) and happens at the back of the nose. ObjectivesThe overall objective of this research is to discuss the prevalence of NPC in selected regions with a focus on Asia Pacific and Malaysia in particular, the categories, severities, symptoms and the existing electronic methods of investigating NPC symptoms. The research also identifies the weaknesses and strengths of the existing methods as inputs for propose improvement. Most part of the study reviewed the current approaches and techniques that are being proposed in other literatures in order to improve the process of investigating NPC. The study critically evaluates these different methods and the problems associated with them. This study provides some review of literatures indicating relationship between cancer milestones configuration and some NPC patient’s attributes. MethodsThis research presents a detailed description of image based NPC, NPC imagery techniques and image types. Next, continue to NPC segmentation, which includes modelling of NPC detection, and subsequently followed by a NPC image classification process. Finally, the research paper discussing the meaning and impact of approaches and the contribution of researchers in this area. The different of NPC symptoms and their basic method of diagnosis were briefly explained. The basic diagnostic procedure is depicted with six procedures where a patient undergoes if he or she is reported with NPC. The six procedures include Trans oral mirror examination, EBV Serology, Circulating EBV DNA in plasma (Plasma DNA), Nasoendoscopy, Biopsy and NP Screen™. FindingThe strengths and weaknesses of the existing methods are identified and identification model for NPC is proposed. The lesson learned from other methods were evaluated and incorporated into proposed model. This study further describes that segmentation, classification and predication of NPC based on many methods Additionally, a literature survey on segmentation, classification and predication of NPC based on real studies has been discussed.

Abstract 3153: A new circulating plasma/serum Epstein-Barr virus (EBV) DNA test by locked nucleic acid (LNA) PCR technique with enhanced sensitivity in diagnosis &amp; monitoring of patients suffering from nasopharyngeal carcinoma (NPC) and other EBV associated malignancies

Abstract Circulating plasma/serum Epstein-Barr virus (EBV) test has been established as routine first line diagnostic test for Nasopharyngeal Carcinoma (NPC). This real time quantitative PCR test is useful in primary diagnosis of NPC as well as monitoring distant metastases of NPC to lung, liver and distant lymph nodes. However, it is still lacking sensitivity in diagnosis of early stage I/II NPC and local relapse in nasopharynx. Detection sensitivity of this PCR test relies on detecting small fragments of circulating EBV DNA shed into the blood stream from NPC tumor lesions which underwent apoptosis/necrosis. Previous literature already showed that the smaller the molecular size of the circulating EBV DNA targets detected, the higher the detection sensitivity. Using Locked Nucleic Acid (LNA) PCR technique, we have designed a new set of LNA primers for PCR amplification of a much smaller EBV gene target (BamH1-W DNA at 46 bp) than the conventional EBV gene target (BamH1-W at 76 bp). Using this LNA marker, we successfully achieved detection sensitivity of 95.7% in 46 NPC patients (44/46 positive) and detection specificity of 98.3% in 59 normal blood screening subjects (58/59 negative). 89.1% (i.e. 41/46) of the NPC patients had higher EBV DNA gene copies per mL in the LNA marker than the conventional marker. Folds of gene copies per mL of increase of LNA marker over those of conventional marker varied from 6 folds up to 3800 folds in 27/46 NPC patients. The results were confirmed in a validation set of 141 NPC patients (113/141) in a multicenter study under the umbrella of NPC Area of Excellence (AoE) program in Hong Kong with 79.6% (113/142) of NPC patients having higher gene copies per mL of LNA marker than the conventional marker initially tested. On longitudinal monitoring of the new LNA marker in 9 NPC patients for a period of 4.9 months to 5.6 months, higher sensitivity of detection was also achieved at disease onset, during the clinical course of disease and at distant metastases (2/9) than the conventional marker. Three patients with Lymphoepithelioma Like Carcinoma of Lung (LELC) and 3 patients with extra nodal nasal NK/T cell lymphoma were monitored from 3 months to 19.3 months. Detection sensitivity was also higher in the new LNA marker in 3/3 LELC and 1/2 NK/T lymphoma with disease progression whereas 1 NK/T lymphoma with remission had LNA marker remained negative. With such enlightening findings, we are expanding the scope of this new test to many more NPC patients as well as other EBV associated malignancies. We are also recruiting patients in embarking on a large scale prospective study. Citation Format: Timothy T.C. Yip, Hazel Y.Y. Kwok, Sharon S.Y. You, Dora L.W. Kwong, Alvin H.W. Fong, W.W. Cheng, Christopher Lai, Anthony C.C. Shek, Victor W.S. Ma, Maria LiLung, Roger K.C. Ngan. A new circulating plasma/serum Epstein-Barr virus (EBV) DNA test by locked nucleic acid (LNA) PCR technique with enhanced sensitivity in diagnosis &amp; monitoring of patients suffering from nasopharyngeal carcinoma (NPC) and other EBV associated malignancies. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3153.

Plasma Epstein-Barr virus DNA as a biomarker for nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is common in southern China and Southeast Asia. Epstein-Barr virus (EBV) infection is an important etiology for NPC, and EBV genome can be detected in almost all tumor tissues of NPC in this region. Plasma EBV DNA, when quantitatively analyzed using real-time polymerase chain reaction (PCR), has been developed as a biomarker for NPC. In this review, the different clinical applications of plasma EBV DNA in the management of NPC, including screening, monitoring, and prognostication, are discussed. In addition, the biological issues of circulating EBV DNA, including the molecular nature and clearance kinetics, are also explored.

Evaluation of plasma Epstein‐Barr virus DNA load to distinguish nasopharyngeal carcinoma patients from healthy high‐risk populations in Southern China

The utility of circulating Epstein-Barr Virus (EBV) DNA as a tumor marker for nasopharyngeal carcinoma (NPC) detection suggests that it might improve the diagnostic performance of anti-EBV antibody markers in NPC screening. In this study, the authors evaluated whether circulating EBV DNA load is capable of distinguishing NPC patients from high-risk individuals who have positive anti-EBV antibodies. In a population-based NPC screening trial in Sihui City and Zhongshan City, Guangdong Province, China, the authors previously identified 862 high-risk participants with 2 screening markers, immunoglobulin A (IgA) antibodies to EBV capsid antigen (VCA/IgA) and nuclear antigen-1 (EBNA1/IgA). In the current study, real-time polymerase chain reaction was used to measure the baseline plasma EBV DNA load among 825 participants (97%). Follow-up was extended to the end of 2011 to evaluate the diagnostic and predictive values of plasma EBV DNA load. By using 0 copies/mL as the cutoff value, plasma EBV DNA had a sensitivity of 86.8% (33 of 38 patients) for NPC detected within the first year of follow-up, yielding a positive predictive value of 30% (33 of 110 participants) and a negative predictive value of 99.3% (696 of 701 participants). The patients who had early stage NPC had lower sensitivity (81.5%; 22 of 27 patients) than those who had advanced NPC (100%; 11 of 11 patients). For the 14 patients who had NPC detected after 1 year of follow-up, only 50% (7 of 14 patients) tested positive for EBV DNA at baseline. The plasma EBV DNA load may improve the accuracy of diagnosing NPC in high-risk individuals, but it appears to have limited value in screening patients who have early stage NPC and predicting NPC development.