Immunobead multiplex RT-PCR detection of carcinoembryonic genes expressing cells in the blood of colorectal cancer patients
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Richard Douard
Abstract
Circulating cell detection using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques has been studied as a new prognostic factor in colorectal cancer patients. With the view of enhancing detection sensitivity, we developed a new multiplex RT-PCR assay for circulating cell detection based on the expression of carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5; formerly CEA) and CEACAM7 (formerly CGM2).
Between November 2002 and December 2003, 45 stage III-IV, 39 stage I-II colorectal cancer patients, 32 non-colorectal cancer patients and 41 healthy individuals were included. Positive selection using HEA-125 immunobeads was applied to blood samples before mRNA extraction, cDNA synthesis and a multiplex CEACAM5/CEACAM7 RT-PCR assay. For both CEACAM5 and CEACAM7, the limit of detection was found to be as low as 1 expressing cell in 10 6 nucleated blood cells.
The multiplex RT-PCR assay was negative for the 41 healthy individuals and the 32 non-colorectal cancer patients. The test was positive in 53/84 (63%) of the colorectal cancer patients for CEACAM5 and/or CEACAM7, whereas 32/84 (38%) were positive for both markers. Colorectal cancer patients were positive for one of the two markers in 80% of cases (36/45) for stage III-IV patients (CEACAM5 73%, CEACAM7 51%) and in 44% of cases (17/39) for stage I-II patients.
This multiplex RT-PCR assay with two markers proved to be more sensitive than use of a single marker in detecting circulating tumour cells. The discrepant expression of CEACAM5 and CEACAM7 may label circulating tumour cells that have different levels of differentiation and subsequent aggressive behaviour.
References
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©2005 by Walter de Gruyter Berlin New York
Articles in the same Issue
- Quality control for SELDI analysis
- Immunobead multiplex RT-PCR detection of carcinoembryonic genes expressing cells in the blood of colorectal cancer patients
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- Whole genome amplification of buccal cell DNA: genotyping concordance before and after multiple displacement amplification
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Articles in the same Issue
- Quality control for SELDI analysis
- Immunobead multiplex RT-PCR detection of carcinoembryonic genes expressing cells in the blood of colorectal cancer patients
- Optimization and evaluation of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) with reversed-phase protein arrays for protein profiling
- Simultaneous determination of HIV antibodies, hepatitis C antibodies, and hepatitis B antigens in dried blood spots –a feasibility study using a multi-analyte immunoassay
- Hematopoietic cytokines in the sera of patients with pancreatic cancer
- Modulation of translation factor's gene expression by histone deacetylase inhibitors in breast cancer cells
- Whole genome amplification of buccal cell DNA: genotyping concordance before and after multiple displacement amplification
- Y-Chromosome short tandem repeat (STR) haplotypes in a Campania population sample
- TaqMan assays for genotyping of single nucleotide polymorphisms present at a disease susceptibility locus on chromosome 6
- The secretion of ibuprofen metabolites interferes with the capillary chromatography of urinary homovanillic acid and 4-hydroxy-3-methoxymandelic acid in neuroblastoma diagnosis
- Selective measurement of HCHO in urine using direct liquid-phase fluorimetric analysis
- A spectrophotometric micromethod for determining erythrocyte protoporphyrin-IX in whole blood or erythrocytes
- Simultaneous analysis of MDR1 C3435T, G2677T/A, and C1236T genotypes by multiplexed mutagenically separated PCR
- Measurement of reticulocyte and red blood cell indices in patients with iron deficiency anemia and β-thalassemia minor
- Chitotriosidase activity in colostrum from African and Caucasian women
- Integration between point-of-care cardiac markers in an emergency/cardiology department and the central laboratory: methodological and preliminary clinical evaluation
- Controlled storage conditions prolong stability of biochemical components in whole blood
- Poor knowledge and faulty thinking regarding hemolysis and potassium elevation
- Delayed effects of short-term transdermal application of 7-oxo-dehydroepiandrosterone on its metabolites, some hormonal steroids and relevant proteohormones in healthy male volunteers
- Evaluation of a novel semi-automated HPLC procedure for whole blood cyclosporin A confirms equivalence to adjusted monoclonal values from Abbott TDx
- Evaluation of the Bio-Rad VARIANT™ II HbA 2/HbA 1C Dual Program for measurement of hemoglobin concentrations and detection of variants
- Interferences in coagulation tests – evaluation of the 570-nm method on the Dade Behring BCS analyser
- No evidence for involvement of the human inducible nitric oxide synthase gene in susceptibility to coronary artery disease
- National survey on the use of measurement of cholinesterase activity in serum
