PURPOSE: To establish a sensitive assay for the specific detection of carcinoembryonic antigen (CEA)-expressing tumor cells in the bone marrow of patients with colorectal cancer and other CEA-positive carcinomas. PATIENTS AND METHODS: A CEA-specific nested reverse transcriptase (RT) polymerase chain reaction (PCR) assay was developed and optimized using limiting dilutions of a CEA-positive cancer cell line mixed with normal bone marrow cell specimens. The optimized test was then used to examine bone marrow samples obtained from 15 patients with abdominal carcinomas (colorectal, n = 10; pancreatic, n = 3; gastric, n = 2) and six patients with breast cancer. Specificity was assessed by examination of 56 negative controls (malignant hematologic disease, n = 28; nonmalignant disease conditions, n = 5; healthy bone marrow donors, n = 8; normal peripheral-blood samples, n = 15). For 11 patients with abdominal carcinomas, immunostaining evaluations were performed using an anti-CEA and an anticytokeratin antibody, and the results compared with the nested PCR assay. RESULTS: In the sensitivity calibration system, single CEA-expressing tumor cells were detected in 2 to 5 x 10(7) normal bone marrow cells. All 56 control samples failed to amplify. This demonstrates that mRNAs coding for highly homologous CEA-related antigens expressed by various lineages of blood cells do not interfere. Bone marrow samples from 10 of 15 patients with abdominal cancers and four of six breast cancer patients scored positive, indicating micrometastatic bone disease. Four of 11 samples from the gastrointestinal cancer patients were found to be positive by the PCR method, but were negative with the immunocytology method. CONCLUSION: Since approximately 30% of the colorectal carcinoma patients that score negative in immunocytology staining of bone marrow samples have been reported to relapse, earlier diagnosis of the presence of malignant cells is needed. Our result that samples scoring positive in the described CEA-specific PCR test remained negative by two immunostaining methods suggests a higher sensitivity. We conclude that PCR amplification of CEA mRNA may lead to an earlier diagnosis of micrometastatic bone disease in patients with CEA-expressing carcinomas.
PURPOSE: To establish a sensitive assay for the specific detection of carcinoembryonic antigen (CEA)-expressing tumor cells in the bone marrow of patients with colorectal cancer and other CEA-positive carcinomas. PATIENTS AND METHODS: A CEA-specific nested reverse transcriptase (RT) polymerase chain reaction (PCR) assay was developed and optimized using limiting dilutions of a CEA-positive cancer cell line mixed with normal bone marrow cell specimens. The optimized test was then used to examine bone marrow samples obtained from 15 patients with abdominal carcinomas (colorectal, n = 10; pancreatic, n = 3; gastric, n = 2) and six patients with breast cancer. Specificity was assessed by examination of 56 negative controls (malignant hematologic disease, n = 28; nonmalignant disease conditions, n = 5; healthy bone marrow donors, n = 8; normal peripheral-blood samples, n = 15). For 11 patients with abdominal carcinomas, immunostaining evaluations were performed using an anti-CEA and an anticytokeratin antibody, and the results compared with the nested PCR assay. RESULTS: In the sensitivity calibration system, single CEA-expressing tumor cells were detected in 2 to 5 x 10(7) normal bone marrow cells. All 56 control samples failed to amplify. This demonstrates that mRNAs coding for highly homologous CEA-related antigens expressed by various lineages of blood cells do not interfere. Bone marrow samples from 10 of 15 patients with abdominal cancers and four of six breast cancerpatients scored positive, indicating micrometastatic bone disease. Four of 11 samples from the gastrointestinal cancerpatients were found to be positive by the PCR method, but were negative with the immunocytology method. CONCLUSION: Since approximately 30% of the colorectal carcinomapatients that score negative in immunocytology staining of bone marrow samples have been reported to relapse, earlier diagnosis of the presence of malignant cells is needed. Our result that samples scoring positive in the described CEA-specific PCR test remained negative by two immunostaining methods suggests a higher sensitivity. We conclude that PCR amplification of CEA mRNA may lead to an earlier diagnosis of micrometastatic bone disease in patients with CEA-expressing carcinomas.
Authors: Ulrich Guller; Paul Zajac; Annelies Schnider; Beatrix Bösch; Stefan Vorburger; Markus Zuber; Giulio Cesare Spagnoli; Daniel Oertli; Robert Maurer; Urs Metzger; Felix Harder; Michael Heberer; Walter Richard Marti Journal: Ann Surg Date: 2002-12 Impact factor: 12.969
Authors: G G Wulf; B Jürgens; T Liersch; W Gatzemeier; H Rauschecker; C Buske; M Hüfner; W Hiddemann; B Wörmann Journal: J Cancer Res Clin Oncol Date: 1997 Impact factor: 4.553
Authors: Barbara K Zehentner; Heather Secrist; Dawn C Hayes; Xinqun Zhang; Richard C Ostenson; Steven Loop; Gary Goodman; Raymond L Houghton; David H Persing Journal: Mol Diagn Ther Date: 2006 Impact factor: 4.074
Authors: Linda W Martin; Jonathan D'Cunha; Xiaofei Wang; Debra Herzan; Lin Gu; Naif Abraham; Todd L Demmy; Frank C Detterbeck; Shawn S Groth; David H Harpole; Mark J Krasna; Kemp Kernstine; Leslie J Kohman; G Alexander Patterson; David J Sugarbaker; Robin T Vollmer; Michael A Maddaus; Robert A Kratzke Journal: J Clin Oncol Date: 2016-02-29 Impact factor: 44.544