A rapid and reliable enzyme immunoassay PCR-based screening method to identify EBV-carrying gastric carcinomas.

Abstract

Epstein-Barr virus (EBV) is associated with a substantial number of gastric adenocarcinomas worldwide, as confirmed by EBER1/2-RNA in situ hybridization (RISH). In the present study, we developed a rapid and sensitive PCR-based prescreening method for the detection of EBV in gastric carcinomas to reduce the amount of laborious EBER1/2-RISH assays to be performed. The method was evaluated by testing gastric adenocarcinomas (n = 242) using both BamHI W PCR-enzyme immunoassay (EIA) and EBER1/2-RISH, in combination with appropriate DNA and RNA quality controls. Seventy-four percent of the paraffin-embedded gastric adenocarcinomas had good DNA quality as shown by beta-globin polymerase chain reaction (PCR) after proteinase K and boiling pretreatment, whereas after DNA purification this was increased to 90%. Thirty-two percent of all cases were EBV-DNA positive after PCR-EIA, whereas 10% of these gastric cancers contained EBV transcripts in the neoplastic cells as confirmed by EBER1/2-RISH. Interestingly, only samples with high optical density (OD) 405/630 values in PCR-EIA, equivalent to the maximum reading of the assay as determined by the positive control, contained EBV-positive tumor cells in the EBER1/2-RISH. In contrast, the weak positive samples, as determined by low OD readings in the PCR-EIA were EBER1/2-RISH negative. In conclusion, high OD values in EBV PCR-EIA are very valuable to prescreen EBV-carrying gastric carcinomas as confirmed by EBER1/2-RISH. Only these samples and those with poor DNA quality will require testing in the EBER1/2-RISH, thereby reducing the amount of laborious RISH assays with 85%.

• Authors: Josine van Beek, Axel zur Hausen, Elma Klein Kranenbarg, Ralph J Warring, Elisabeth Bloemena, Mikael E Craanen, Cornelis J H van de Velde, Jaap M Middeldorp, Chris J L M Meijer, Adriaan J C van den Brule
• Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc (Mod Pathol) Vol. 15 Issue 8 Pg. 870-7 (Aug 2002) ISSN: 0893-3952 [Print] United States
• PMID: 12181273 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Actins
• DNA, Viral
• RNA, Messenger
• RNA-Binding Proteins
• Ribosomal Proteins
• RPL22 protein, human

Topics

• Actins (analysis)
• Adenocarcinoma (diagnosis, virology)
• DNA, Viral (analysis)
• Epstein-Barr Virus Infections (virology)
• Herpesvirus 4, Human (isolation & purification)
• Humans
• Immunoenzyme Techniques (methods)
• Immunohistochemistry
• In Situ Hybridization
• Polymerase Chain Reaction (methods)
• RNA, Messenger (analysis)
• RNA-Binding Proteins (analysis)
• Ribosomal Proteins
• Sensitivity and Specificity
• Stomach Neoplasms (diagnosis, virology)
• Tumor Virus Infections (virology)