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Assessment of the methods for the detection of Epstein-Barr virus nucleic acids and related gene products in Hodgkin's disease.
Lab Invest. 1993 Oct; 69(4):483-90.LI

Abstract

BACKGROUND

Epstein-Barr virus (EBV) is present in the pathogenic cells of a significant number of cases of Hodgkin's disease, particularly of the mixed cellularity subtype. EBV remains latent and the incidence of detection rate of the genomes and gene products varies greatly with the methods employed.

EXPERIMENTAL DESIGN

From a pool of 137 cases of Hodgkin's disease previously studied by cold in situ hybridization (ISH) for the presence or absence of EBV DNA and the immunohistochemical reactivity with anti-latent membrane protein 1 antibody, we selected 24 cases (12 EBV DNA-positive, 12 EBV DNA-negative) for Southern blotting, as well as for study with nonisotopic EBER and BHLF1 oligonucleotide probes and amplification of DNA by polymerase chain reaction. EBV positive cases were further tested with anti-ZEBRA (BZLF1) antibody.

RESULTS

The EBV DNA detection rate was found to be lower with Southern blotting compared to ISH and IHC methods because 8 of the 12 EBV positive cases were positive with BamHI W probe and only 7 (of the 8) with XhoI 1.9 kb probe. These 7 cases contained monoclonal episomal circular EBV genomes. All EBV DNA-positive cases showed EBER gene transcription by ISH. EBER probes also reacted with small lymphocytes in all EBV DNA+ and 9 EBV DNA- cases. These EBER-positive small lymphocytes were detected neither by BamHI W DNA probe nor by immunohistochemical methods with anti-latent membrane protein 1 and anti-EBNA2 antibodies. Polymerase chain reaction produced a positive result in all EBV DNA+ cases and 2 (of the 9) EBV DNA- cases containing EBER+ small lymphocytes. This discrepancy was attributed to amplification of EBV in reactive lymphocytes. Anti-ZEBRA antibody was positive in 2 cases (one BHLF1+) suggesting infrequent viral replication and probable abortive lytic cycle.

CONCLUSIONS

ISH and immunohistochemical methods are more sensitive than Southern blot for detecting EBV in Hodgkin and Reed-Sternberg cells of Hodgkin's disease. Polymerase chain reaction appears to be very sensitive but is less sensitive and specific (amplification of EBV DNA of non-neoplastic lymphocytes) than ISH with non-isotopic EBER oligoprobes, which often detects and localizes EBV in pathogenic cells even when they are in small numbers and also in a few small lymphocytes. In addition, this method offers the advantage of being applied to routinely processed tissue sections.

Authors+Show Affiliations

Anatomical Pathology Laboratory, Centre Hospitalier Universitaire de Purpan, Toulouse, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8231115

Citation

Brousset, P, et al. "Assessment of the Methods for the Detection of Epstein-Barr Virus Nucleic Acids and Related Gene Products in Hodgkin's Disease." Laboratory Investigation; a Journal of Technical Methods and Pathology, vol. 69, no. 4, 1993, pp. 483-90.
Brousset P, Meggetto F, Chittal S, et al. Assessment of the methods for the detection of Epstein-Barr virus nucleic acids and related gene products in Hodgkin's disease. Lab Invest. 1993;69(4):483-90.
Brousset, P., Meggetto, F., Chittal, S., Bibeau, F., Arnaud, J., Rubin, B., & Delsol, G. (1993). Assessment of the methods for the detection of Epstein-Barr virus nucleic acids and related gene products in Hodgkin's disease. Laboratory Investigation; a Journal of Technical Methods and Pathology, 69(4), 483-90.
Brousset P, et al. Assessment of the Methods for the Detection of Epstein-Barr Virus Nucleic Acids and Related Gene Products in Hodgkin's Disease. Lab Invest. 1993;69(4):483-90. PubMed PMID: 8231115.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Assessment of the methods for the detection of Epstein-Barr virus nucleic acids and related gene products in Hodgkin's disease. AU - Brousset,P, AU - Meggetto,F, AU - Chittal,S, AU - Bibeau,F, AU - Arnaud,J, AU - Rubin,B, AU - Delsol,G, PY - 1993/10/1/pubmed PY - 1993/10/1/medline PY - 1993/10/1/entrez SP - 483 EP - 90 JF - Laboratory investigation; a journal of technical methods and pathology JO - Lab. Invest. VL - 69 IS - 4 N2 - BACKGROUND: Epstein-Barr virus (EBV) is present in the pathogenic cells of a significant number of cases of Hodgkin's disease, particularly of the mixed cellularity subtype. EBV remains latent and the incidence of detection rate of the genomes and gene products varies greatly with the methods employed. EXPERIMENTAL DESIGN: From a pool of 137 cases of Hodgkin's disease previously studied by cold in situ hybridization (ISH) for the presence or absence of EBV DNA and the immunohistochemical reactivity with anti-latent membrane protein 1 antibody, we selected 24 cases (12 EBV DNA-positive, 12 EBV DNA-negative) for Southern blotting, as well as for study with nonisotopic EBER and BHLF1 oligonucleotide probes and amplification of DNA by polymerase chain reaction. EBV positive cases were further tested with anti-ZEBRA (BZLF1) antibody. RESULTS: The EBV DNA detection rate was found to be lower with Southern blotting compared to ISH and IHC methods because 8 of the 12 EBV positive cases were positive with BamHI W probe and only 7 (of the 8) with XhoI 1.9 kb probe. These 7 cases contained monoclonal episomal circular EBV genomes. All EBV DNA-positive cases showed EBER gene transcription by ISH. EBER probes also reacted with small lymphocytes in all EBV DNA+ and 9 EBV DNA- cases. These EBER-positive small lymphocytes were detected neither by BamHI W DNA probe nor by immunohistochemical methods with anti-latent membrane protein 1 and anti-EBNA2 antibodies. Polymerase chain reaction produced a positive result in all EBV DNA+ cases and 2 (of the 9) EBV DNA- cases containing EBER+ small lymphocytes. This discrepancy was attributed to amplification of EBV in reactive lymphocytes. Anti-ZEBRA antibody was positive in 2 cases (one BHLF1+) suggesting infrequent viral replication and probable abortive lytic cycle. CONCLUSIONS: ISH and immunohistochemical methods are more sensitive than Southern blot for detecting EBV in Hodgkin and Reed-Sternberg cells of Hodgkin's disease. Polymerase chain reaction appears to be very sensitive but is less sensitive and specific (amplification of EBV DNA of non-neoplastic lymphocytes) than ISH with non-isotopic EBER oligoprobes, which often detects and localizes EBV in pathogenic cells even when they are in small numbers and also in a few small lymphocytes. In addition, this method offers the advantage of being applied to routinely processed tissue sections. SN - 0023-6837 UR - https://www.unboundmedicine.com/medline/citation/8231115/Assessment_of_the_methods_for_the_detection_of_Epstein_Barr_virus_nucleic_acids_and_related_gene_products_in_Hodgkin's_disease_ L2 - https://medlineplus.gov/hodgkindisease.html DB - PRIME DP - Unbound Medicine ER -