Related Experiment Video
Updated: May 20, 2026
Isolation and Quantification of Epstein-Barr Virus from the P3HR1 Cell Line
Published on: September 28, 2022
Quantification of Epstein-Barr virus DNA is helpful for evaluation of chronic active Epstein-Barr virus infection
Yuichi Sakamoto1, Yasushi Mariya, Kohmei Kubo
1Department of Laboratory Medicine, Aomori Prefectural Central Hospital, Aomori, Japan. 97036ys@jichi.ac.jp
Insights
Monitoring Epstein-Barr virus (EBV) DNA levels using real-time PCR in peripheral blood offers a noninvasive method to track chronic active EBV infection (CAEBV) and associated complications like EBV-HLH. This approach aids in understanding disease progression and treatment response.
Area of Science:
- Virology
- Immunology
- Molecular Diagnostics
Background:
- Chronic active Epstein-Barr virus infection (CAEBV) can manifest with diverse symptoms, including fever and liver dysfunction.
- Diagnosis of EBV-associated diseases, such as EBV-hemophagocytic lymphohistiocytosis (EBV-HLH) and EBV-positive lymphoma, often relies on invasive bone marrow analysis.
- Cytomegalovirus (CMV) poses significant risks, particularly pneumonia and retinitis, in immunocompromised individuals.
Observation:
- A 30-year-old Japanese female patient presented with recurrent fever, fatigue, and liver dysfunction indicative of CAEBV and acute hepatitis.
- Liver biopsy confirmed EBV presence, and peripheral blood EBV DNA levels were significantly elevated (>1.0 × 10⁷ copies/mL).
- Real-time PCR for EBV and CMV DNA in peripheral blood provided a noninvasive means to monitor viral activity and clinical status.
Findings:
- Quantifying EBV DNA levels via real-time PCR effectively reflected disease activity and response to immunosuppressive and antiviral therapies.
- Following bone marrow transplantation for EBV-HLH, rising EBV DNA levels correlated with lymphocyte increases, indicating potential relapse.
- Simultaneous detection of CMV DNA using PCR proved more sensitive than the standard CMV antigenemia test.
Implications:
- Real-time quantitative PCR for EBV DNA is a valuable tool for noninvasively assessing the clinical course of CAEBV and related conditions.
- This method aids in timely therapeutic adjustments and monitoring disease progression in immunocompromised patients.
- Close monitoring of viral DNA levels can improve patient management and potentially outcomes in EBV-associated diseases.
Abstract:
Chronic active Epstein-Barr virus infection (CAEBV) presents with chronic or recurrent infectious mononucleosis-like symptoms, such as low-grade fever, liver dysfunction, lymphadenopathy, and hepatosplenomegaly. Immunological methods are useful for the diagnosis of viral infections. However, CAEBV patients do not necessarily have high titers of Epstein-Barr virus (EBV)-specific antibodies. Hosts that are immunocompromised after hematopoietic stem cell transplantations sometimes suffer from systemic EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) and EBV-positive lymphoma. Patients with EBV-associated diseases are often diagnosed by analyses of bone marrow. Cytomegalovirus (CMV) can cause serious pneumonia or retinitis in immunocompromised hosts. In order to noninvasively understand the clinical status of patients with EBV-associated diseases, we conducted real-time polymerase chain reaction (PCR) methods in their peripheral blood in order to quantify EBV and CMV DNA levels, which reflect viral activity. Here, we describe a 30-year-old Japanese female patient with CAEBV. The patient had repeated fever, fatigue, and liver dysfunction. The histopathological results of liver biopsies were positive for EBV-encoded RNA-1. Acute hepatitis was associated with the EBV infection. The whole-blood EBV DNA levels were high and above 1.0 × 10⁷ copies/mL. After immunosuppressive and antiviral therapies, EBV DNA levels lowered. However, she had to receive bone marrow transplantation because of her EBV-HLH. As the number of lymphocytes increased in the post-transplantation period, EBV DNA levels gradually increased again. The simultaneous detection of CMV DNA was more sensitive than the CMV antigenemia test that is often used to diagnose CMV infections. Unfortunately, the patient died due to a fungal infection. Observing EBV DNA levels closely with real-time quantitative PCR methods is helpful for evaluating the changes in the clinical course.
