Abstract
Introduction: Epstein-Barr virus (EBV) may serve as a target in therapeutic treatments, thus reliable diagnostic results are necessary. Objective: The aim of this study was to evaluate the accuracy of EBV detection by in situ hybridization (ISH) using five commercial probes in formalin-fixed and paraffin-embedded samples of nodular sclerosis Hodgkin's lymphoma (HL), and to compare the results with immunohistochemistry (IHC) and polymerase chain reaction (PCR). Material and method: Thirty samples were selected, 28 were lymph nodes, one bone marrow and one mediastinum. The following parameters were analyzed: signal intensity; proportionality of positive cells; quality of the reaction according to comfort for evaluation, sign quality and homogeneity of labeled cells; background reaction; morphology; presence of artifacts; and positivity in other non-neoplastic cells. All samples were analyzed for EBV detection using the five probes, IHC for latent membrane protein type 1 (LMP1) and PCR for Epstein Barr virus nuclear antigen 1 (EBNA1). Statistical analyses were performed with the R1 software; Fleiss' test and Cohen Kappa index of 5% were considered significant. Results: The detection by IHC-LMP1 was 26.7% (8/30) and 66.7% (20/30) by PCR-EBNA1. All probes detected EBV. Positivity was observed in 42/90 (46.7%), 38/90 (42.2%), 45/90 (50%), 27/90 (30%) and 61/90 (67.8%) for probes A, B, C, D and E, respectively. Discussion: All five probes demonstrated positivity. Conclusion: Probe E showed better rate (67.8%), sensitivity, specificity and accuracy (100%), a very good correlation among the different observers and with PCR, besides great cost-benefits relation.
Highlights
Epstein-Barr virus (EBV) may serve as a target in therapeutic treatments, reliable diagnostic results are necessary
The aim of this study was to evaluate the accuracy of EBV detection by in situ hybridization (ISH), using five different commercially available probes, in São Paulo, Brazil, in cases of HLNS, and compare it to the results of IHC and polymerase chain reaction (PCR)
All probes used for ISH-EBER detected the EBV (Figure)
Summary
Epstein-Barr virus (EBV) may serve as a target in therapeutic treatments, reliable diagnostic results are necessary. All samples were analyzed for EBV detection using the five probes, IHC for latent membrane protein type 1 (LMP1) and PCR for Epstein Barr virus nuclear antigen 1 (EBNA1). Epidemiological studies have estimated that 90%-95% of the human adult population are infected with EBV[1,2,3,4,5,6] and has been transmitted intermittently through saliva In most individuals, this virus promotes an asymptomatic to subclinical infection[3, 6] and persists latently in the host. EBV is a linear, double-stranded deoxyribonucleic acid (DNA) virus with 172,000 base pairs, which can remain latent in the lymphoid cells in its episomal form[3, 5] These cells express genes, among others, a small non-polyadenylated ribonucleic acid (RNA) chain that does not translate a protein, consisting of two fragments known as EBER1 (166 nucleotides) and EBER2 (172 nucleotides).
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