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- Title
A Rapid and Reliable Enzyme Immunoassay PCR-Based Screening Method to Identify EBV-Carrying Gastric Carcinomas.
- Authors
van Beek, Josine; zur Hausen, Axel; Kranenbarg, Elma Klein; Warring, Ralph J.; Bloemena, Elisabeth; Craanen, Mikael E.; van de Velde, Cornelis J.H.; Middeldorp, Jaap M.; Meijer, Chris J.L.M.; van den Brule, Adriaan J.C.
- Abstract
Epstein-Barr virus (EBV) is associated with a substantial number of gastric adenocarcinomas world wide, as confirmed by EBER1/2-RNA in situ hybridization (RISH). In the present study, we develop 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 β-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%.
- Subjects
GASTRIC diseases; ADENOCARCINOMA; EPSTEIN-Barr virus; ENZYME-linked immunosorbent assay; SOLID-phase analysis; CANCER cells
- Publication
Modern Pathology, 2002, Vol 15, Issue 8, p870
- ISSN
0893-3952
- Publication type
Article
- DOI
10.1097/01.MP.0000024147.43288.B1