scholarly journals Detection of Epstein-Barr virus in autoimmune and Thalassemia patients using new Immunoblot assay

2015 ◽  
Vol 9 (1) ◽  
pp. 13-16
Author(s):  
Alaa Younis Mahdy Al-Hamadany Al-Hamadany ◽  
Basima A. Abdullah Abdullah

A new immunoblot assay, composed of five Epstein-Barr virus (EBV)-encoded recombinantproteins virus capsid antigen [VCA] gp125, p19, p22, early antigen [EA], and EBNA-1 IgG, wasused to manifest the EBV infection and look at the antibody pattern to EBV proteins in the serumof both autoimmune disorders and Thalassemia patients and compare the observations with thosein normal healthy controls. Serum samples from 35 rheumatoid arthritis patients, 20 SLE, 20autoimmune hypothyroid diseases, 35 Thalassemia patients and 20 healthy controls were testedfor EBV IgG antibodies by an immunoblot assay (Euroline). The results showed that the highpercentage recorded was 50% in acute infection. Followed by 30% at late infection, while latephase with loss EBNA-1 and reactivated infection were 10% compared to the normal healthycontrols. Our study showed an increased EBV activation among the autoimmune patient groupscompared to the normal healthy controls. Further studies are required to delineate the associationbetween the etiology of autoimmune disorders and EBV.

1999 ◽  
Vol 37 (8) ◽  
pp. 2709-2714 ◽  
Author(s):  
M. Buisson ◽  
B. Fleurent ◽  
M. Mak ◽  
P. Morand ◽  
L. Chan ◽  
...  

A new immunoblot assay, composed of four Epstein-Barr virus (EBV)-encoded recombinant proteins (virus capsid antigen [VCA] p23, early antigen [EA] p138, EA p54, and EBNA-1 p72), was compared with an immunofluorescence assay on a total of 291 sera. The test was accurate in 94.5% of cases of primary EBV infection, while an immunoglobulin G anti-VCA p23 band with strong intensity correlated with reactivation.


Author(s):  
R. Stephens ◽  
K. Traul ◽  
D. Woolf ◽  
P. Gaudreau

A number of antigens have been found associated with persistent EBV infections of lymphoblastoid cells. Identification and localization of these antigens were principally by immunofluorescence (IF) techniques using sera from patients with nasopharyngeal carcinoma (NPC), Burkitt lymphoma (BL), and infectious mononucleosis (IM). Our study was mainly with three of the EBV related antigens, a) virus capsid antigen (VCA), b) membrane antigen (MA), and c) early antigens (EA) using immunoperoxidase (IP) techniques with electron microscopy (EM) to elucidate the sites of reactivity with EBV and EBV infected cells.Prior to labeling with horseradish peroxidase (HRP), sera from NPC, IM, and BL cases were characterized for various reactivities by the indirect IF technique. Modifications of the direct IP procedure described by Shabo and the indirect IP procedure of Leduc were made to enhance penetration of the cells and preservation of antigen reactivity.


2021 ◽  
pp. 1-6
Author(s):  
Alev Çetin Duran ◽  
Tuğba Kula Ati

Objective: Epstein-Barr virus (EBV) can cause different clinical pictures from infectious mononucleosis (IM) to malignancies such as B-cell lymphomas, Burkitt’s lymphoma, nasopharyngeal carcinoma, and Hodgkin lymphoma. VCA-IgM, VCA-IgG, EBNA-1 IgG antibodies are the most commonly used antibodies in revealing the serological profile. This study aimed to examine the serological profiles of patients with suspected EBV infection and to interpret the atypical profiles encountered. Methods: The results of VCA-IgM, VCA-IgG, and EBNA-1 IgG antibodies studied in the Microbiology Laboratory between 2017-2019 were evaluated retrospectively. EBV serological tests (VCA-IgM, VCA-IgG, and EBNA-1 IgG) were performed according to the manufacturer’s recommendations using the chemiluminescent microparticle immunoassay (CMIA) method (Architect, Abbott, Wiesbaden, Germany). Results: Of the 2486 patients evaluated, 1341 (53.9%) were male, 1145 (46.1%) were female, and the average age was determined as 16.93 ± 19.5. EBV past infection was detected in 56.65% of the cases, the acute infection was detected in 17.25%, and 21.09% did not encounter EBV. Atypical serological profile was detected in 5.01%. As an atypical profile, the most common positivity of three antibodies together (3.90%), then isolated VCA-IgG positivity (0.91%), and isolated EBNA-1 IgG positivity (0.20%) were determined. It was determined that 24.24% of the cases with an atypical profile were immunosuppressive patient.  Conclusions: The rate of encountering EBV in our study is 78.91%. The atypical EBV serological profile rate was found to be 5.01%. Approximately one-fourth of the cases with an atypical profile was found to be in the patient group with immune disorders. It is thought that antibody tests are not sufficient to determine the stage of infection, especially in these patient groups, and further tests should be performed. It has been demonstrated that serological monitoring is required for the interpretation of atypical profiles. Keywords: Epstein-Barr virüs, serological tests, chemiluminescent microparticle immunoassay (CMIA), atypical serological profile


Blood ◽  
2001 ◽  
Vol 97 (5) ◽  
pp. 1165-1171 ◽  
Author(s):  
Servi J. C. Stevens ◽  
Erik A. M. Verschuuren ◽  
Inge Pronk ◽  
Wim van der Bij ◽  
Martin C. Harmsen ◽  
...  

Posttransplant lymphoproliferative disease (PTLD) is a frequent and severe Epstein-Barr virus (EBV)–associated complication in transplantation recipients that is caused by iatrogenic suppression of T-cell function. The diagnostic value of weekly EBV DNA load monitoring was investigated in prospectively collected unfractionated whole blood and serum samples of lung transplantation (LTx) recipients with and without PTLD. In PTLD patients, 78% of tested whole blood samples were above the cut-off value of quantitative competitive polymerase chain reaction (Q-PCR) (greater than 2000 EBV DNA copies per mL blood), with the majority of patients having high viral loads before and at PTLD diagnosis. Especially in a primary EBV-infected patient and in patients with conversion of immunosuppressive treatment, rapid increases in peripheral blood EBV DNA load diagnosed and predicted PTLD. In non-PTLD transplantation recipients, only 3.4% of the whole blood samples was above the cut-off value (P < .0001) despite heavy immune suppression and cytomegalovirus (CMV)-related disease. These findings illustrate the clinical importance of frequent EBV DNA load monitoring in LTx recipients. The increased EBV DNA loads in PTLD patients were restricted to the cellular blood compartment, as parallel serum samples were all below cut-off value, which indicates absence of lytic viral replication. EBV+ cells in PTLD patients have a very short doubling time, which can be as low as 56 hours, thereby creating the need for high screening frequency in high-risk patients. Furthermore, it is shown that EBV and CMV can reactivate independently in LTx recipients and that EBV DNA load monitoring may be useful in discriminating PTLD from rejection.


2019 ◽  
Vol 7 (12) ◽  
pp. 628 ◽  
Author(s):  
Takuya Shuto ◽  
Jun Nishikawa ◽  
Kanami Shimokuri ◽  
Ayaka Yanagi ◽  
Tatsuya Takagi ◽  
...  

Background: Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is classified as one of the molecular subtypes of gastric cancer. We used droplet digital polymerase chain reaction (ddPCR) to enable highly sensitive and quantitative detection of EBV. Methods: EBV-DNA load was calculated based on the copy number of the BamH1-W fragment of EBV by ddPCR, and the cut-off value of EBV-DNA load was set. We conducted both ddPCR and EBER1 ISH to examine whether their results coincided in 158 gastric cancer specimens of unknown EBV status. We prepared 26 biopsy specimens and 49 serum samples including EBVaGC and assayed them by ddPCR. Results: The median values of EBV-DNA load for EBVaGC and EBV-negative control were 17.0 and 0.00308, respectively. A cut-off value of 0.032 was determined for which the sensitivity was 1. Among the 158 gastric cancer specimens, 14 lesions were judged as EBV-positive by the 0.032 cut-off value determined by ddPCR. The results of ddPCR and EBER1 ISH were in complete agreement. Even when using a biopsy specimen as a sample for ddPCR, the EBV-DNA load of all EBVaGCs was larger than the cut-off value. Conclusions: We established a new method of diagnosing EBVaGC from tissue samples by ddPCR.


Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 487
Author(s):  
Peter A. C. Maple ◽  
Bruno Gran ◽  
Radu Tanasescu ◽  
David I. Pritchard ◽  
Cris S. Constantinescu

Background: Epstein–Barr virus (EBV) infection is strongly associated with multiple sclerosis (MS). Helminth infection can downregulate antiviral immune responses, potentially protecting against MS, but with a theoretical risk for reactivating latent EBV infection. Objective: To investigate parameters of EBV infection and their relationship with disease activity in people with MS (PwMS) therapeutically vaccinated with Necator americanus (hookworm). Methods: Sequential serum samples from 51 PwMS; 26 therapeutically infected (25 larvae) with N. americanus and 25 controls were tested for EBV virus capsid antigen (VCA) IgG and IgM, EBV nuclear antigen-1 (EBNA-1) IgG, and EBV early antigen (EA) IgG. Disease activity was assessed by periodic MRI. Significance was set at p < 0.05. Results: All PwMS were EBV VCA IgG and EBNA-1 IgG positive, and 35.2% were EBV EA IgG positive. EBV antibody levels were generally stable, and EBV reactivation in PwMS was not demonstrated by significant increases in IgG titre over 12 months. Disease activity was most frequent in PwMS possessing high levels of EBV VCA IgG (>600 units/mL) or EBNA-1 IgG (>150 units/mL); however, there was no association with hookworm treatment. Interpretation: Therapeutic hookworm vaccination was not associated with EBV reactivation. Multiple sclerosis disease activity was associated with high levels of EBV VCA IgG or EBNA-1 IgG.


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