Development of a sensitive, specific reverse transcriptase polymerase chain reaction-based assay for epithelial tumour cells in effusions

Citation
M. Sakaguchi et al., Development of a sensitive, specific reverse transcriptase polymerase chain reaction-based assay for epithelial tumour cells in effusions, BR J CANC, 79(3-4), 1999, pp. 416-422
Citations number
30
Categorie Soggetti
Oncology,"Onconogenesis & Cancer Research
Journal title
BRITISH JOURNAL OF CANCER
ISSN journal
00070920 → ACNP
Volume
79
Issue
3-4
Year of publication
1999
Pages
416 - 422
Database
ISI
SICI code
0007-0920(199902)79:3-4<416:DOASSR>2.0.ZU;2-V
Abstract
We developed a sensitive and specific method for the detection of epithelia l cancer cells in effusions with a two-stage molecular-based assay which co mbined enrichment for cancer cells by immunomagnetic bead selection and rev erse transcriptase polymerase chain reaction (RT-PCR) detection of epitheli al glycoprotein 2 (EGP-2) RNA. Preliminary experiments indicated that immun obead selection was essential to avoid occasional false-positive RT-PCR res ults, and this method detected ten breast cancer cells electively added to 10(7) cytologically negative effusion cells. We studied 110 cases of pleura l (n = 68) and peritoneal (n = 42) effusions (30 from patients with known c arcinoma and 80 from those without known carcinoma), and the results were c ompared with cytological findings. Of 18 effusions that were cytologically positive or suspicious for malignant cells, 17 (94%) were positive far EGP- 2 RNA (the one negative sample was from a patient who recently received com bination chemotherapy). Of 92 cytologically negative samples, 11 (12%) were positive for EGP-2, including six patients with a history of previous or c urrent carcinoma. Our method appears to be highly specific and increases th e sensitivity of detection of malignant cells; it may be a useful adjunct t o routine cytopathological examination.