The application of flow cytometric DNA analysis in detecting the presence of malignant cells in ovarian carcinoma peritoneal fluids

Abstract

Objective To compare flow cytometric detection of malignant cells with standard cytological evaluation in patients with ovarian carcinoma. Setting The City Hospital Trust, The Women's Hospital and CRC Trials Unit, Birmingham. Subjects Forty‐three patients with histologically proven ovarian carcinoma and positive cytology, and a control population of 20 patients undergoing surgery for benign gynaecological conditions. Methods Prospective, blinded study examining ascitic fluid or peritoneal washings obtained at primary surgery by flow cytometric DNA analysis and cytological examination. Results Flow cytometry detected aneuploid cells in 27/43 (63%) of malignant and 7/20 (35%) of benign fluid specimens. In malignant samples the mean aneuploid count was 38.5% (range 1–98%) with a mean S‐phase fraction of 5.2% (range 0–33.9%). In benign specimens the mean aneuploid count was 30.4% (range 14.5–66.4%). Based on these results, the overall sensitivity of cytometric detection of malignant cells was 714Y0, specificity 65%, with a positive predictive value of 85.1%. False positivity was found mainly in patients with benign ovarian cysts. Further examination revealed four false negative and four false positive results, where the peritoneal fluid and ovarian tissue DNA ploidy status concurred. Assuming such results to be correct increased the sensitivity of the test to 88.5YO and specificity to 85%. Conclusions Although flow cytometry can glean information beyond the capabilities of cytological assessment, using the premise that aneuploid cells alone indicate malignancy, it remains secondary to cytology in the detection of malignant cells in peritoneal fluids.

Evaluation of flow cytometric immunophenotyping and DNA analysis for detection of malignant cells in serosal cavity fluids

“…15,40 Nevertheless, the majority of published studies did not perform that comparison. 16,42,43 On the other hand, some authors did not explore the DI average and just refer to the FCM results in terms of aneuploid or diploid. [44][45][46][47][48] We also observed that DI shows no difference in effusions due to infiltration of malignant epithelial cells or hematopoitic malignancy or due to hepatocellular carcinoma developing in cirrhotic liver.…”

“…13,14 Some reported that DNA flow cytometry was in general less sensitive than cytology for the detection of malignant cells and a higher percentage of false-positive cases were seen by FCM. 15,16 Others reported increasing sensitivity and specificity. 17 Diagnostic algorithms were developed based on tumor marker measurements in effusions.…”

Diagnostic Value of p53 Protein and Flow Cytometric DNA Analysis in the Study of Serous Effusions

“…FCM DNA analysis has been shown to be useful for the detection of malignant cells in a variety of serous effusions. [13][14][15]23,24,27,28,32 The criteria for the determination of aneuploidy differs from author to author. We consider as aneuploidy only those cases with a clearly separate extra peak separable from the diploid cell population.…”

“…This is similar to that reported in other effusion studies. [13][14][15]23,24,27,28,32 In all these studies, the major dilemma in applying FCM to detecting malignant cells is the premise that an absence of aneuploid cells indicates absence of malignancy. All tumors can contain diploid-only cells, 4,10 and aneuploidy is not always synonymous with malignancy.…”

Heparanase expression: a potential ancillary diagnostic tool for distinguishing between malignant cells and reactive mesothelium in body cavity effusions

“…Cells from reactive or hyperplastic mesothelium shed from body cavity surfaces in various biological settings may present a wide range of deviation from normal cellular morphology, making it difficult, or even impossible, to distinguish them from malignant cells by means of purely cytological criteria. Many diagnostic procedures, such as DNA analysis by means of flow or image cytometry, [1][2][3] AgNOR evaluation, 4,5 in situ hybridization, 6,7 identification of K-ras mutations by PCR, 8 and detection of human telomerase in cells from body fluids, 9,10 have been investigated as possible ancillary diagnostic means for meeting the challenge, but none of these has been widely accepted as a plausible diagnostic procedure. Likewise, application of various monoclonal antibodies aimed at discriminating between the different cell types is, as of now, of limited value.…”