Efficacy of Quantitative Analysis of Epstein-Barr Virus-Infected Peripheral Blood Lymphocytes by in Situ Hybridization of Eber1 After Living-Related Liver Transplantation

Nakazawa, Yūichi; Chisuwa, Hisanao; Ikegami, Toshihiko; Matsunami, Hidetoshi; Ichikawa, Tetsuo; Oh‐ishi, Tsutomu; Kawasaki, Seiji

doi:10.1097/00007890-199705150-00029

Cited by 9 publications

(4 citation statements)

References 12 publications

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“…LPD was suspected in patients with lymphadenopathy, pulmonary infiltration, gastrointestinal tract bleeding, or unexplained allograft dysfunction. The diagnosis of LPD was established by pathology or the detection of EBER by the ISH assay (18,33). IM was diagnosed by clinical findings and serological examinations as follows: positive for anti-viral capsid antigen (VCA) immunoglobulin G (IgG) and/or IgM and negative for anti-EB nuclear antigen (EBNA) antibody.…”

Section: Methodsmentioning

confidence: 99%

“…A biopsy is invasive and labor-intensive, so several methods to detect the virus load in the peripheral blood for the early diagnosis of LPD and CAEBV have been developed. Spontaneous outgrowth of EBV-infected B cells in vitro (27,30), in situ hybridization (ISH) using EBV-encoded small RNA (EBER) probe (18), and quantitative PCR assays are now used to determine the EBV load in peripheral blood mononuclear cells (PBMNC) (2,26,27,29). Previously, we reported that quantitative PCR was useful for diagnosing and monitoring EBV infections (37).…”

mentioning

confidence: 99%

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Quantitative Analysis of Epstein-Barr Virus Load by Using a Real-Time PCR Assay

et al. 1999

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To measure the virus load in patients with symptomatic Epstein-Barr virus (EBV) infections, we used a real-time PCR assay to quantify the amount of EBV DNA in blood. The real-time PCR assay could detect from 2 to over 107 copies of EBV DNA with a wide linear range. We estimated the virus load in peripheral blood mononuclear cells (PBMNC) from patients with symptomatic EBV infections. The mean EBV-DNA copy number in the PBMNC was 103.7 copies/μg of DNA in patients with EBV-related lymphoproliferative disorders, 104.1 copies/μg of DNA in patients with chronic active EBV infections, and 102.2copies/μg of DNA in patients with infectious mononucleosis. These numbers were significantly larger than those in either posttransplant patients or immunocompetent control patients without EBV-related diseases. In a patient with infectious mononucleosis, the virus load decreased as the symptoms resolved. The copy number of EBV DNA in PBMNC from symptomatic EBV infections was correlated with the EBV-positive cell number determined by the in situ hybridization assay (r = 0.842; P < 0.0001). These results indicate that the real-time PCR assay is useful for diagnosing symptomatic EBV infection and for monitoring the virus load.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Quantitative Analysis of Epstein-Barr Virus Load by Using a Real-Time PCR Assay

et al. 1999

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“…EBER-1, one of the EBERs, is detectable in virtually all EBV-infected cells and is expressed at very high levels, reaching 10 7 molecules per cell, although no protein is apparently translated. Previous studies have used in situ hybridisation with an EBER-1 probe to detect and count EBV-infected cells [30,31]. This technique is widely used to detect EBV in tissue specimens [32].…”

Section: Technical Aspects In Measuring Ebv Load In Peripheral Bloodmentioning

confidence: 99%

Measuring Epstein–Barr virus (EBV) load: the significance and application for each EBV‐associated disease

Kimura

Ito

Suzuki

et al. 2008

Reviews in Medical Virology

137

142

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Because Epstein-Barr virus (EBV) is ubiquitous and persists latently in lymphocytes, simply detecting EBV is insufficient to diagnose EBV-associated diseases. Therefore, measuring the EBV load is necessary to diagnose EBVassociated diseases and to explore EBV pathogenesis. Due to the diverse biology of EBV, the significance of measuring EBV DNA and the optimal type of specimen differ among EBV-associated diseases. Recent advances in molecular technology have enabled the EBV genome to be quantitated rapidly and accurately. Real-time polymerase chain reaction (PCR) is a rapid and reliable method to quantify DNA and is widely used not only as a diagnostic tool, but also as a management tool for EBV-associated diseases. However, each laboratory currently measures EBV load with its own ''homebrew'' system, and there is no consensus on sample type, sample preparation protocol, or assay units. The EBV real-time PCR assay system must be standardised for large-scale studies and international comparisons.

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“…Data from the Riddler study indicated that using semiquantitative polymerase chain reaction (PCR), patients with PTLD had a viral load greater than 5000 EBV genome copies/10 6 PBMC (27). Other studies confirmed these findings (28)(29)(30)(31)(32)(33). An association between PTLD and EBV detection in plasma has also been reported (33).…”

Section: Detection Of Ebv Nucleic Acids or Protein In Tissuementioning

confidence: 95%

Epstein-Barr Virus Infection in Transplant Recipients: Sumary of a Workshop on Surveillance, Prevention and Treatment

Allen

Alfieri

Preiksaitis

et al. 2002

Canadian Journal of Infectious Diseases

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Diseases caused by the Epstein-Barr virus are of great significance among organ transplant recipients. One of these diseases, post-transplant lymphoproliferative disease, is a major complication among organ transplant recipients. Management of this entity is problematic due to the difficulties with laboratory surveillance, diagnosis, prevention and treatment. A group of Canadian and American experts was assembled to discuss these aspects of Epstein-Barr virus diseases in Canadian organ transplant recipients. This report summarizes the relevant background literature and levels of evidence in relation to the outcomes of the deliberations and recommendations by the expert panel.

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Efficacy of Quantitative Analysis of Epstein-Barr Virus-Infected Peripheral Blood Lymphocytes by in Situ Hybridization of Eber1 After Living-Related Liver Transplantation

Cited by 9 publications

References 12 publications

Quantitative Analysis of Epstein-Barr Virus Load by Using a Real-Time PCR Assay

Quantitative Analysis of Epstein-Barr Virus Load by Using a Real-Time PCR Assay

Measuring Epstein–Barr virus (EBV) load: the significance and application for each EBV‐associated disease

Epstein-Barr Virus Infection in Transplant Recipients: Sumary of a Workshop on Surveillance, Prevention and Treatment

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