title: Real-time reverse transcription-polymerase chain reaction assay for GA733-2 mRNA in the detection of metastatic carcinoma cells in serous effusions.
abstract: Cytomorphologic and immunocytologic examination alone provide only limited sensitivity for the detection of metastatic carcinoma cells in many cases of serous effusions. The specificity of conventional reverse transcription-polymerase chain reaction (RT-PCR) methods for detection of epithelial gene transcripts is low owing to the ectopic expression of many such genes in nonepithelial cells. For the detection of metastatic carcinoma cells, we describe a quantitative real-time RT-PCR assay for GA733-2 messenger RNA encoding an epithelial glycoprotein (EGP-2) that binds to the monoclonal antibody BerEP4. With serial dilutions of BerEP4-positive SK-BR-3 breast carcinoma cells, the RT-PCR assay was able to detect 10 carcinoma cells in a background of 10(6) lymphoid cells compared with 10(2) to 10(3) carcinoma cells detectable by immunocytologic examination. We analyzed 51 serous effusions, including 25 malignant metastatic effusions, by the real-time RT-PCR assay. Receiver operating characteristic curve analyses demonstrated a sensitivity of 96% corresponding to a specificity of 98% for a correct diagnosis of metastatic effusions. These results provide evidence that the GA733-2 real-time RT-PCR assay is a specific and sensitive tool for the detection of metastasis of BerEP4-positive primary tumors in serous effusion specimens.
authors.affiliation: Department of Cytopathology, Georg August University, Goettingen, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Nagel
authors.forename: Carola
authors.initials: C
authors.lastname: Werner
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hemmerlein
mesh_terms: D000951:Antigens, Neoplasm / Q000235:genetics*; D015815:Cell Adhesion Molecules / Q000235:genetics*; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D006801:Humans; D008297:Male; D009362:Neoplasm Metastasis; D016066:Pleural Effusion, Malignant / Q000175:diagnosis*; D012333:RNA, Messenger / Q000032:analysis*; D012372:ROC Curve; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; D012333:RNA, Messenger