Detection of Epstein- Barr virus infection in lymphoma: ELISA and PCR method

Abstract

Background: Latent Epstein- Barr virus (EBV) genomes are found in the malignant cells of approximately one-third of Hodgkin's lymphoma (HL) cases. Detection of EBV viral DNA could potentially be used as a biomarker of disease activity. Our goal was to compare of EBV DNA detection in samples obtained from lymphoma patients versus controls. Methods: One milliliter uncoagulated and 1ml coagulated blood sample for DNA extraction and serum analysis using ELISA for IgG anti EBNA-1 were obtained from 44 lymphoma patients and from 44 normal controls, respectively. EBV genome, EBNA-2, was examined from DNA extracts of paraffin embedded and blood samples using Nested PCR with type specific inner primers. Results: Positive results for ELISA, Blood and biopsy PCR in study group were, 84.1%, 27.3% and 13.6%, respectively. However, these results in control group were 47.7% and 16% for ELISA and Blood PCR assays, respectively. Positive results in ELISA, Blood PCR and Biopsy PCR in Hodgkin and non-Hodgkin patients were found in 21(84%), 6(24%), 4(16%) and 16(84.2%), 6(31.6%), 2(10.5%) of specimens, respectively. No significant differences in EBV detection were found between these two patient groups (p values for ELISA, Blood PCR and Biopsy PCR were 0.26, 0.73 and 0.68, respectively). Conclusion: Comparison of ELISA and Blood PCR results in children and adult patients with the same age of controls have showed difference in ELISA results of children, only. None of the test results have showed statistically significant difference between Hodgkin and non-Hodgkin patients. However, the mean of ELISA results in Hodgkin patients was higher as compared with controls. Blood PCR assay cannot be recommended as a biomarker of disease activity in EBV positive Hodgkin's lymphoma patients.