Urinary Carcinoembryonic Antigen Research Articles

A dye-sensitized solar cell acting as the electrical reading box of an immunosensor: Application to CEA determination

Monitoring cancer biomarkers in biological fluids has become a key tool for disease diagnosis, which should be of easy access anywhere in the world. The possibility of reducing basic requirements in the field of electrochemical biosensing may open doors in this direction. This work proposes for this purpose an innovative electrochemical immunosensing system using a photovoltaic cell as an electrical reading box. Immunosensing ensures accuracy, the electrochemical-ground of the device ensures sensitivity and detectability, and the photovoltaic cell drives the system towards electrical autonomy. As proof-of-concept, Carcinoembryonic antigen (CEA) was used herein, a cancer biomarker of clinical relevance.In brief, a conductive glass with a fluorine doped tin oxide film was used as conductive support and modified with anti-CEA by means of a bottom-up approach. All stages involved in the biochemical modification of the FTO surface were followed by electrochemical techniques, namely electrochemical impedance spectroscopy and cyclic voltammetry. This electrode acted as counter electrode of a dye-sensitized solar cells, and the electrical output of this cell was monitored for the different concentrations of CEA. Under optimized conditions, the device displayed a linear behaviour against CEA concentration, from 5 pg/mL to 15 ng/mL. The immunosensor was applied to the analysis of CEA in urine from healthy individual and spiked with the antigen.Overall, the presented approach demonstrates that photovoltaic cells may be employed as an electrical reading box of electrochemical biosensors, yielding a new direction towards autonomous electrochemical biosensing.

Abstract 5136: Bladder cancer diagnostic algorithms in hematuria populations.

Abstract Queens University Belfast, the Belfast Health Trust and Randox Laboratories are working together to create risk and diagnostic classifiers for patients with haematuria. Biomarker profiles may fail to differentiate between all possible causes of hematuria, but it would be a huge advancement if benign and malignant causes were differentiated. Our objective was to determine whether single biomarkers or multivariate algorithms could significantly improve on the predictive power of an algorithm based on demographics for detection of urothelial cancer in patients presenting with hematuria. We collected demographic and clinical information from 157 patients (80 urothelial cancers and 77 with confounding pathologies) who had presented with hematuria and were recruited to a case control study. For each patient we measured 22 urinary biomarkers and serum Carcinoembryonic antigen (CEA) using either ELISAs or the Randox biochip technology, BAT. We analysed the collated data, using Forward Wald binary logistic regression analyses. Algorithms based on demographic variables were designated as Prior Predicted Probability (PPP). We then created algorithms incorporating PPP as a single variable, together with biomarkers.We determined Areas under the Receiver Operating Characteristic (AUROC) for differentially expressed single biomarkers and multivariate algorithms. AUROC for CEA, Bladder Tumour Antigen (BTA) and Nuclear matrix protein (NMP22) = 0.74; 0.74; and 0.79, respectively. Vascular endothelial growth factor, dDimer, Fas, hyaluronidase, interleukin(IL) IL-1α, IL-6 and IL-8 were also differentially expressed (T-test; p < 0.05). PPP included age and smoking years. In combination, PPP, NMP22 and Epidermal Growth Factor (EGF) achieved AUROC = 0.90, significantly improving upon the AUROC for PPP which was 0.76. Although neutrophil-associated gelatinase lipocalin (NGAL) levels were similar in urines from UC and control patients, a multivariate classifier which comprised age, serum CEA and urinary NGAL and interleukin 8 achieved a respectable AUROC = 0.844. Biomarkers significantly improved upon the AUROC statistic based on demographics. Despite non differential urinary levels across UC and controls, NGAL made a significant contribution to a diagnostic classifier exemplifying the unpredictable nature of the complexities involved in collectives of biomarkers. The impact of biomarkers in multivariate algorithms for bladder cancer diagnosis in patients with hematuria. Abogunrin F, O'Kane HF, Ruddock MW, Stevenson M, Reid CN, O'Sullivan JM, Anderson NH, O'Rourke D, Duggan B, Lamont JV, Boyd RE, Hamilton P, Nambirajan T, Williamson KE. Cancer. May 15;118(10) [2012] American Cancer Society Inc Citation Format: Michael Stevenson, Cherith Reid, Brian Duggan, Mark Ruddock, Kate E. Williamson. Bladder cancer diagnostic algorithms in hematuria populations. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5136. doi:10.1158/1538-7445.AM2013-5136

Urine carcinoembryonic antigen determination in urinary bladder bilharziasis predicts carcinoma in patients with premalignant lesions: Observation of 43 cases in Egypt

Both urinary bilharziasis and urothelial neoplasia are associated with increased production of tissue carcinoembryonic antigen (CEA). Urine and serum CEA were determined in 43 patients with urinary bladder carcinoma including 22 post bilharzial and 21 non-biharzial cases, in addition to 10 normal control cases. A significant increase was detected in both urine and serum CEA levels with bladder carcinoma compared to control cases. Urinary CEA was significantly elevated in 86% of bilharzial versus 62% in nonbilharzial bladder carcinoma. Only 10.5% of control cases had urinary CEA elevation. The mean urinary CEA in bilharzial was higher than that of nonbilharzial carcinoma, but the difference was not statistically significant. There was a definite relationship between urine CEA and the stage of malignancy; the higher the stage, the higher the level of urine CEA. No relationship could be detected between the stage of malignancy and serum CEA, or between the grades of malignancy and urine or serum CEA levels. In conclusion, urinary CEA is more useful than serum CEA in the early detection of bilharziasis-associated urothelial carcinoma.

Carbon Nanoparticle-Enhanced Immunoelectrochemical Detection for Protein Tumor Marker with Cadmium Sulfide Biotracers

We have developed a sensitive electrochemical immunoassay system for the detection of a protein tumor marker, carcinoembryonic antigen (CEA), that is based on a carbon nanoparticle (CNP)/poly(ethylene imine) (PEI)-modified screen-printed graphite electrode (CNP-PEI/SPGE) covered with anti-CEA antibodies. The signal amplification strategy--using CdS nanocrystals as biotracers and CNPs to enhance electron transfer--improves the sensitivity and detection limit for CEA, suggesting that this system holds promise for development into a point-of-care or disposable home-care self-diagnostic tool. This biosensor is based on a sandwich complex immunoassay, which we assembled from sequential layers of the anti-CEA antibody (alphaCEA) on CNP-PEI/SPGE, the CEA sample, and the CdS nanocrystal quantum dots (QDs) sensitized with alphaCEA (alphaCEA-CdS QD). We used square wave anodic stripping voltammetry (SWASV) to amplify the signal current response obtained from the dissolved alphaCEA-CdS QDs. The calibration curve for CEA concentration was linear in the range of 0.032-10 ng/mL; the detection limit (estimated as the mean of the blank sample plus three times the standard deviation obtained on the blank sample) was 32 pg/mL (equivalent to 160 fg in a 5 microL sample). This method is suitably precise and sensitive to function as a means of determining urinary CEA, which is a better marker than serum CEA for the early detection of urothelial carcinoma.

Urine carcinoembryonic antigen levels are more useful than serum levels for early detection of Bilharzial and non-Bilharzial urinary bladder carcinoma: observations of 43 Egyptian cases

Background Both urinary bilharziasis and urothelial neoplasia are associated with increased production of tissue carcinoembryonic antigen (CEA).

Utility of Slot‐Blot‐ELISA as a New, Fast, and Sensitive Immunoassay for Detection of Carcinoembryonic Antigen in the Urine Samples of Patients with Various Gastrointestinal Malignancies

Carcinoembryonic antigen (CEA) is the most widely used clinical tumor marker. CEA immunoassay has found acceptance as a diagnostic adjunct in clinical diagnosis of gastrointestinal tumors (GIT). Several immunoassays have been established for detection of CEA in plasma, serum, tissue, feces, and urine of cancer patients using polyclonal or monoclonal antibodies raised against CEA. Some of these assays display both high sensitivity and specificity for the detection of CEA. However, these assays require special and highly expensive equipment and the procedures require long periods for their completion. In the present study, we established a Slot‐Blot Enzyme Linked Immunosorbent Assay (SB‐ELISA), based on anti‐CEA monoclonal antibody (CEA‐mAb), as a new, simple, fast, cheap, and non‐invasive immunodiagnostic technique for detection of CEA in the urine of GIT patients. Urine and serum samples were collected from 248 GIT patients (58 with pancreatic cancer, 20 with hepatoma, 23 with ampullary carcinoma, 15 with hilar cholangiocarcinoma, 28 with gastric cancer, 14 with esophageal cancer, and 90 with colorectal cancer). Moreover, urine and serum samples were collected from 50 healthy individuals to serve as negative controls. The traditional ELISA technique was used for determination of CEA in the sera of GIT patients using anti‐CEA monoclonal antibody. A comparison between the results of both techniques (ELISA and SB‐ELISA) was carried out. The traditional ELISA detected CEA in the sera of 154 out of 248 GIT patients with a sensitivity of 59.8%, 51.7% positive predictive value (PPV) and 75.37% negative predictive value (NPV). In addition, it identified 15 false positive cases out of 50 healthy individuals with a specificity of 70%. The urinary CEA was identified by a Western blotting technique and CEA‐mAb at a molecular mass of 180 Kda. The developed SB‐ELISA showed higher sensitivity, specificity, PPV, and NPV (70.1%, 78%, 62.4%, and 82.13%, respectively) for detection of CEA in the urine of GIT patients. The semi‐quantitative SB‐ELISA showed a higher overall efficiency of 72.8% versus 63.4% in the case of the quantitative ELISA, for detection of CEA. In conclusion, SB‐ELISA is more efficient for detection of CEA in gastrointestinal tumors. It is a simple, rapid, non‐invasive, and sensitive assay. Moreover, all steps of the SB‐ELISA are performed at room temperature, without the use of expensive equipment; this may enhance the application of this assay in field studies and mass screening programs.

Urine carcinoembryonic antigen levels are more useful than serum levels for early detection of Bilharzial and non-Bilharzial urinary bladder carcinoma: Observations of 43 Egyptian cases

BackgroundBoth urinary bilharziasis and urothelial neoplasia are associated with increased production of tissue carcinoembryonic antigen (CEA).Patients and methodsUrine and serum CEA were determined in 43 patients with urinary bladder carcinoma including 22 post bilharzial and 21 nonbiharzial cases, in addition to 10 normal control cases.ResultsA significant increase was detected in both urine and serum CEA levels with bladder carcinoma compared to control cases. Urinary CEA was significantly elevated in 86% of bilharzial, versus 62% in nonbilharzial bladder carcinoma. Only 10.5% of control cases had urinary CEA elevation. The mean urinary CEA in bilharzial, was higher than that of nonbilharzial carcinoma, but the difference was not statistically significant. There was a definite relationship between urine CEA and the stage of malignancy; the higher the stage, the higher the level of urine CEA. No relationship could be detected between the stage of malignancy and serum CEA, or between the grades of malignancy and urine or serum CEA levels.ConclusionUrinary CEA is more useful than serum CEA in the early detection of urotherlial carcinoma particularly if provoked by bilharziasis. Its level is also correlated with the tumor stage.

Increased serum carcinoembryonic antigen level in patients undergoing colon neobladder replacement compared with ileal neobladder replacement

Objectives. To compare the serum and urinary carcinoembryonic antigen (CEA) levels for assessment of possible risk of malignant transformation in patients with orthotopic neobladder. Methods. The serum and urinary levels of CEA, nutritional status, and acid-base and electrolyte balances were studied in 87 patients after radical cystectomy (22 with ileal neobladder, 28 with colon neobladder, and 37 with ileal conduit). The results of these groups were compared. Results. The serum CEA level in patients with colon neobladder, ileal neobladder, and ileal conduit was 5.4 ± 3.0, 3.7 ± 1.6, and 3.1 ± 1.5 ng/mL, respectively. The serum CEA level in the colon neobladder group was significantly higher than the levels in the remaining two groups ( P <0.05); 16 patients (57%) with colon neobladder had elevated serum CEA values (ie, greater than 5 mg/mL). Elevated serum CEA was observed in only 5 (23%) and 3 (8%) patients with ileal neobladder or ileal conduit, respectively. The serum CEA value in these patients was associated with the urinary CEA value ( P <0.001), but not the other factors examined. Conclusions. These findings suggest that colon bladder replacement caused significantly increased serum CEA values compared with ileal neobladder or ileal conduit; however, the elevated serum CEA level correlated with the urinary CEA level, irrespective of other clinical factors. Therefore, the elevated serum CEA in the colon neobladder group may have been due to reabsorption of CEA in urine rather than to an association with malignant changes in the bowel segments used for neobladder creation.

Elevated tumour markers in patients with Balkan endemic nephropathy.

Balkan nephropathy (BEN) is commonly associated with urothelial cancer. Urothelial cancer is manifested in the advanced stage of disease. The aim of this study was to facilitate early detection of urothelial cancer in BEN patients and their family members living in an endemic region by using tumour markers, carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA), and a putative marker, ferritin. Fifteen BEN patients with normal renal function, 17 with renal failure (BEN-RF), 13 healthy members of their families (HFM), 14 patients with glomerulonephritis (GN) and 12 healthy controls (C) were studied. Serum CEA levels in BEN patients were within normal limits, however, in BEN-RF patients they were significantly increased over HFM (p<0.05). Serum TPA levels in BEN and BEN-RF patients were significantly higher than in the C and HFM groups (p<0.05). Urinary CEA was not significantly different between the groups studied. Urinary TPA levels in HFM (median 125 U/l, BEN (236 U/l) and BEN-RF (275 U/l) were significantly increased over C (30 U/l), however, TPA levels were increased also in GN patients (437 U/l). None of the BEN patients studied developed urothelial cancer during the ten years' follow-up. Markedly elevated urinary TPA-like levels in all patients studied (HFM, BEN, BEN-RF, GN) suggest that urinary TPA may not be a reliable tumour marker. However, the clinical relevance of high TPA levels in BEN patients should be evaluated.

Simultaneous determination of urinary CEA, ferritin and TPA in Egyptian bladder cancer patients.

Urinary carcinoembryonic antigen (CEA), ferritin (Fer) and tissue polypeptide antigen (TPA) were determined in 328 cases (106 with bladder cancer, 152 with non-malignant urinary tract disease and 70 healthy controls). CEA was determined by the kit supplied by Roche Diagnostica (CEA EIA Doumab 60), ferritin by the Tandem-E Fer kit supplied by Hybritech and TPA by the Prolifigen TPA-IRMA kit supplied by Sangtec Medical. The results of this work revealed that combined determination of urine CEA and Fer, CEA and TPA or Fer and TPA showed higher sensitivity than determination of the individual markers. There was no significant difference between combined and individual marker determination with respect to false positivity in non-malignant urinary tract diseases. At 97% specificity, the sensitivities of urine CEA, Fer and TPA were 82.1%, 71.7% and 90.6%, respectively, while combined urine CEA & Fer, CEA & TPA and Fer & TPA showed sensitivities of 92.5%, 99.1% and 98.1%, respectively. When the specificity was related to the entire non-cancer group (patients with benign urinary tract diseases and normal controls), some reduction in the sensitivities of the combined markers was noted compared to the normal group only. In conclusion, combined determination of urine markers is superior to determination of individual markers in the diagnosis of bladder cancer.

Immunological and biochemical characterization of urinary carcinoembryonic antigen and its clinical significance

The immunobiochemical properties of urinary carcinoembryonic antigen (U-CEA) purified from non-tumor-bearing patients with urinary tract infection were investigated. Moreover, immunohistochemical localization of CEA in the normal urothelium was studied and clinical significance of U-CEA was also analyzed statistically. The purified U-CEA showed a molecular weight of approximately 160,000 by the Western blotting and an antigenicity identical to the purified CEA (Calbiochem Corp.). Amino acids analysis of U-CEA revealed nearly the same values as the purified CEA or the theoretical values of CEA, except for several amino acids susceptible to hydrolysis and sugar chains. Therefore, it was assumed that the U-CEA was a new CEA-related antigen, different from nonspecific cross-reacting antigen (NCA) or even CEA itself. Immunohistochemically, CEA was found mainly at the cytoplasmic membrane in normal urothelial cells. The urine samples, collected from patients with various urological disorders, were divided into the inflammation group, showing 5 leukocytes/hpf or more in the urine, and the non-inflammation group, showing less than 5 leukocytes/hpf. The groups were subdivided and U-CEA levels were statistically studied; it was found that the U-CEA levels strongly reflected the presence of inflammation, rather than the presence of urothelial tumors. Consequently, it was considered that CEA was present in the normal urothelium and that destruction and regeneration of these cells due to tumor or inflammation might release CEA, not NCA, into the urine.

Evaluation of a Double-Monoclonal Radioimmunoassay for the Measurement of Carcinoembryonic Antigen in the Urine of Patients with Bladder Cancer

A double-monoclonal radioimmunoassay (RIA) has been used to detect urinary carcinoembryonic antigen (CEA) in 204 patients attending for diagnostic cystoscopy. Elevated levels of urinary CEA were found in 50% of patients with bladder tumours, and these levels correlated with both the grade and stage of the tumour. The sensitivity proved to be 58% and the specificity 74%, which compares with previous studies using polyclonal RIA kits. As a monitoring test, urinary CEA proves to be as useful as urine cytology. The use of a more specific assay does not increase the clinical value of the test which still lacks the sensitivity and specificity needed to allow it to be used to monitor recurrence of low grade tumours (Ta, Tis and T1).

Possibilities for an extended classification of bladder cancer.

Patients with bladder cancer were evaluated for T-class, histo-pathological grade and U-CEA (urinary carcinoembryonic antigen) before treatment and the cytological picture 4 months after treatment. Previous work has shown that these variables are not significantly intercorrelated. Scores were computed, consisting of the sums of these (dichotomized) variables. In a statistical analysis with the life-table technique, the scored variables have been log-rank tested for a prognostic trend. In 155 patients, p for symptom-free survival between subgroups with low and high scores was 0.0019 and for relative survival 0.0005. This implies that a combination of variables may have predictive value in bladder cancer.

The value of urinary cytology, serum and urinary carcinoembryonic antigen, rheumatoid factors, and urinary immunoglobulin concentration as tumor markers or prognostic factors in predicting progression of superficial bladder cancer.

We followed 67 patients with superficial bladder cancer (TA, T1) for 12-48 months (mean, 26.8 months) after surgical removal of the tumors. Every 3-4 months, when the patients came for cystoscopic control, we evaluated serum and urinary carcinoembryonic antigen (CEA), urinary cytology, serum rheumatoid factors, and urinary IgG, IgA, and IgM concentrations. The purpose was to look for markers or prognostic factors in patients with superficial bladder carcinoma other than characteristics of the excised tumors themselves. We emphasized data that can be obtained on patient follow-up. Our results were correlated with tumor recurrence and tumor progression rates. In contrast with other reports, only urinary immunoglobulins (especially IgG) proved to have prognostic value. However, urinary CEA and (even more so) urinary cytology are fairly good tumor markers: they were positive when the bladder tumors were present and, in the case of recurrence, before recurrent tumors were visible.

Prediction of survival and recurrence in bladder carcinoma

We have followed a large population of patients receiving radiation treatment for bladder carcinoma with respect to survival and recurrence-free survival. Bivariate and multivariate life table analyses have been performed using a set of independent variables. The most important were T class, grade (G), urinary carcinoembryonic antigen (U-CEA) taken before treatment and cytological analysis 4 months after treatment. We compared the usual way of classifying a patient (T + G) with the combination of U-CEA and cytology since the latter two variables seemed to have great prognostic importance. The analyses show that T + G gives the best significance for survival (p = 0.0003) while U-CEA and cytology is better for recurrence-free survival (rho = 0.0002). 0.0002).

Changes in the Urine Protein Profile Following Intravesical Doxorubicin

Proteinuria was studied before and after 1 instillation of intravesical doxorubicin in 35 patients with superficial bladder cancer, 25 of whom had primary or secondary carcinoma in situ. An increased plasma protein transudation was demonstrated across the bladder wall in patients without microscopic hematuria. The levels of urinary carcinoembryonic antigen indicated that its release into the urine after doxorubicin treatment for T1 tumors is associated with a successful response.

Carcinoembryonic antigen localization in benign and malignant transitional epithelium.

Transitional epithelium from 30 cases, including transitional cell carcinoma of various grades and stages, carcinoma in situ, cystitis cystica, and glandularis, as well as normal bladder epithelium, was studied for carcinoembryonic antigen (CEA) content by the immunoperoxidase technique. The results indicate that CEA is present in normal transitional epithelium and that its level frequently decreases in transitional cell carcinoma. These results are related to the findings in studies of urinary CEA excretion.

Assessment of serial CEA determinations in urine of patients with bladder carcinoma.

Carcinoembryonic antigen (CEA) levels in urine and serum from 294 patients with bladder cancer in varying stages have been clinically evaluated. All urine samples were obtained from patients with intact renal function and without bacterial infection in the bladder. The samples were collected before, during, and at follow-up examination after radiotherapy. They were perchloric acid extracted before being assayed in a double-antibody radioimmunoassay. The geometric mean of urine CEA levels for patients with primary tumors of Stage T1 or T2 was significantly lower than that for those with Stage T3 or T4 disease. The urine CEA levels for patients with tumors of various histologic grades did not differ. The urine CEA levels decreased from before to after radiation treatment of the primary tumor. Patients with recurrence within six months after undergoing primary treatment had higher initial mean urine CEA levels than did those without evidence of recurrence. The prognostic information for recurrence was limited to the more advanced tumors. Differences were also found between the means of samples taken before recurrence and after treatment of recurrent tumors; with regression of the tumor, a lower mean urine CEA level was found; with progression, a higher value. Urine CEA levels before any treatment were higher when the patients had a short survival time. Serum CEA levels were not related to stage or grade of the primary bladder tumor but levels were slightly elevated with metastases. The determination of urine CEA levels seems to be useful in the follow-up of patients with bladder carcinoma because when initially high, it adds to the information of the T classification and predicts early recurrence, and the monitoring of individual patients after primary treatment is useful for detecting recurrence.

Screening tests for detection of bladder cancer

Eighty-one individuals had urine cytologic studies, urinary carcinoembryonic antigen (CEA) determinations, and urinary immunoglobulin levels performed to evaluate various screening tests for the diagnosis of bladder cancer. Urinary cytologic studies detect the presence of bladder malignancy, and while false positive tests were troublesome, remain the primary screening test for bladder cancer. Urinary CEA was of little value in detecting vesical malignancy. Urinary immunoglobulins, particularly IGG and IGA, were significantly elevated in the presence of bladder cancer.

Inverted papilloma of the urinary bladder: ultrastructural and immunologic studies.

The histologic, ultrastructural, and immunologic features of an inverted urinary bladder papilloma were studied with thin-section and freeze-fracture electron microscopy, a specific red blood cell adherence test, and through analysis of urine carcinoembryonic antigen (CEA). The increased number of gap junctions and desmosomes, the presence of thickened and reduplicated basal laminae, and the normal CEA levels found in association with this neoplasm, correlate well with its generally predictable benign behavior.