Evaluation of seven tumour markers in pleural fluid for the diagnosis of malignant effusions

Carcinoembryonic antigen (CEA), carbohydrate antigens 15-3, 19-9 and 72-4 (CA 15-3, CA 19-9 and CA 72-4), cytokeratin 19 fragments (CYFRA 21-1), neuron-specific enolase (NSE) and squamous cell carcinoma antigen (SCC) were evaluated in pleural fluid for the diagnosis of malignant effusions. With a specificity of 99%, determined in a series of 121 benign effusions, the best individual diagnostic sensitivities in the whole series of 215 malignant effusions or in the subgroup of adenocarcinomas were observed with CEA, CA 15-3 and CA 72-4. As expected, a high sensitivity was obtained with SCC in squamous cell carcinomas and with NSE in small-cell lung carcinomas. CYFRA and/or CA 15-3 were frequently increased in mesotheliomas. Discriminant analysis showed that the optimal combination for diagnosis of non-lymphomatous malignant effusions was CEA + CA 15-3 + CYFRA + NSE: sensitivity of 94.4% with an overall specificity of 95%. In malignant effusions with a negative cytology, 83.9% were diagnosed using this association. The association CYFRA + NSE + SCC was able to discriminate adenocarcinomas from small-cell lung cancers. Regarding their sensitivity and their complementarity, CEA, CA 15-3, CYFRA 21-1, NSE and SCC appear to be very useful to improve the diagnosis of malignant pleural effusions.     

肿瘤标志物联合检测在良恶性胸腔积液鉴别诊断中的价值

 目的　探讨胸腔积液4种肿瘤标志物联合检测在良恶性胸腔积液鉴别诊断中的价值。方法　采用电化学发光免疫法检测126例胸腔积液患者（其中恶性组52例,良性组74例）癌胚抗原（CEA）、糖类抗原125（CA125）、糖类抗原15-3（CA15-3）和细胞角蛋白片段19（CYFRA21-1）水平, 并计算上述指标单独和与CEA联合检测在诊断中的敏感度、特异度、准确度和约登指数（YI）。结果　恶性组4种肿瘤标志物水平均明显高于良性组（P＜0.01）。单项检测各种肿瘤标志物的敏感度以CA125最高（90.4 %）,特异度以CYFRA21-1最高（79.7 %）,诊断准确度以CEA和CYFRA21-1最高（71.4 %）,YI以CEA最高（0.41）。联合检测较单项检测敏感度、准确度和YI明显提高,其中CEA、CYFRA21-1和CA15-3三项联合效果最好,敏感度为92.3 %,特异度为78.4 %,准确度为84.1 %,YI值最高为0.71。四项联合敏感度为94.2 %,特异度为75.7 %,准确度为83.3 %,YI值为0.70,与三项联合结果相比差异无统计学意义（P＞0.05）。结论　单项检测的诊断价值有限,CEA、CYFRA21-1和CA15-3三项联合效果最好、最经济,可指导患者恰当选择进一步的侵入性检查手段。     

Diagnostic utility of CYFRA 21-1, carcinoembryonic antigen, CA 125, neuron specific enolase, and squamous cell antigen level determinations in the serum and pleural fluid of patients with pleural effusions.

BACKGROUND: To the authors' knowledge the role of tumor marker determination in the differential diagnosis of pleural effusions has not been established definitively. The current article reports the results of a study of CYFRA 21-1, carcinoembryonic antigen (CEA), cancer antigen 125 (CA 125), squamous cell antigen (SCC), and neuron specific enolase (NSE) in the serum and pleural fluid of patients with pleural effusions of diverse etiologies. METHODS: One hundred forty-six patients with pleural effusions (43 malignant, 47 tuberculous, 32 miscellaneous benign, and 24 paramalignant) were studied prospectively. Levels of CYFRA 21-1, CA 125, CEA, NSE, and SCC were measured by radioimmunoassay in the pleural fluid in all patients and in the serum in 118 patients. RESULTS: There were no significant differences between the serum and pleural fluid levels of tumor markers with the exception of CA 125, which was higher in the pleural fluid. With maximum specificity, the highest sensitivity in the diagnosis of pleural malignancy was obtained with a combination of CYFRA 21-1 (with a cutoff value of 150 U/L), CEA (with a cutoff value of 40 ng/mL), and CA 125 (with a cutoff value of 1000 ng/mL) in pleural fluid. NSE and SCC added no diagnostic value. The simultaneous use of tumor markers and cytology in pleural fluid increased the sensitivity from 55.8% to 81%. CONCLUSIONS: These findings suggest that a combination of CYFRA 21-1, CEA, and CA 125 in the pleural fluid can be a useful addition to pleural cytology in the diagnosis of malignant pleural effusion.     

Diagnostic value of CEA, CA 15-3, CA 19-9, CYFRA 21-1, NSE and TSA assay in pleural effusions

The aim of this study was to evaluate the individual and combined diagnostic utility of six tumor markers in patients with pleural effusion. Pleural and serum levels of carcinoembryonic antigen (CEA), carbohydrate antigen 15-3 (CA 15-3), carbohydrate antigen 19-9 (CA 19-9), cytokeratin fragment 19 (CYFRA 21-1), neuron-specific enolase (NSE) and total sialic acid (TSA) were assayed in 74 patients with pleural effusions (44 malignant and 30 benign). All tumor markers except TSA and NSE were increased in both serum and pleural fluid of patients with malignant diseases. Using the cut-off values 3 ng/ml, 14 U/ml, 5 U/ml, 8 ng/ml and 70 mg/dl for pleural fluid CEA, CA 15-3, CA 19-9, CYFRA 21-1 and TSA, respectively, the sensitivity (%) and specificity (%) of these tumor markers were as follows: CEA; 52/77, CA 15-3; 80/93, CA 19-9; 36/83, CYFRA 21-1; 91/90, TSA; 80/67, for differentiating malignant effusions from benign. When CA 15-3 and CYFRA 21-1 combined, the sensitivity and specificity were increased (100 and 83%, respectively). Classifying the malignant effusions as bronchial carcinoma and malignant pleural mesothelioma, CEA was shown to have the highest sensitivity and specificity (88 and 90%, respectively) while the combination of CEA with other tumor markers increased sensitivity but decreased specificity. According to our results, tumor markers are not suitable for the differential diagnosis of malignancy.     

DNA倍体联合肿瘤标志物检测在胸腔积液诊断中的应用

目的探讨DNA倍体分析联合肿瘤标志物在良、恶性胸腔积液诊断中的价值。方法将108例胸腔积液分为恶性组（68例）和良性组（40例）。除常规细胞学检查外,以流式细胞术（flowcytometry,FCM）检测患者胸腔积液中的DNA倍体,采用化学发光法测定胸腔积液中CEA、CA199、NSE、CYFRA211、SCC、CA125等肿瘤标志物含量。比较DNA倍体联合肿瘤标志物诊断与细胞学诊断的优劣。结果DNA倍体诊断恶性胸腔积液的敏感性、特异性分别为70．6％、95．0％,Youden’s指数为0．656,敏感度稍高于细胞学诊断的65．1％,差异无统计学意义（P〉0．05）。除NSE外,其他5种肿瘤标志物在恶性胸腔积液中浓度均高于良性,差异有统计学意义（P〈0．05）。CYFRA211、CEA、CAl99、CAl25、SCC、NSE的AUC分别为：0．893,0．828,0．759,0．691,0．524及0．490;COV分别为：149．2ng／mL,53．6ng／mL,78．2IU／mL,1559．0IU／mL,48．72ng／mL及78．3ng／mL;敏感性分别为：44．1％,44．1％,35．3％,29．4％,13．2％,5．9％,特异性均为100％。4种肿瘤标志物联合检测＋DNA倍体检测的敏感性为88．2％（60／68）,特异性95％,显著高于细胞学诊断。结论DNA倍体联合CEA、CA199、CYFRA211和CA125检测诊断恶性胸腔积液有较高敏感性,具有定量、快速、价廉、易标准化的特点,且操作简单。     

Evaluation of cytokeratin 19 fragment (CYFRA 21-1) as a tumor marker in malignant pleural effusion

BACKGROUND: The aim was to investigate the diagnostic utility of CYFRA 21-1 (cytokeratin 19 fragment) as a tumor marker in pleural effusion and evaluate the value of combining CYFRA 21-1 and carcinoembryonic antigen (CEA) assays as a diagnostic aid in the malignant pleural effusion. METHODS: One hundred and twenty-six patients (72 malignant and 54 benign pleural effusion) were included in this retrospective study. The effusion levels of CYFRA 21-1 and CEA were measured using radioimmunometric assay. RESULTS: The median values of CYFRA 21-1 in benign and malignant pleural effusion are 15 and 70 ng/ml, respectively. Using a cut-off value of 50 ng/ml, defined at 94% specificity, the diagnostic sensitivity of CYFRA 21-1 for non-small cell lung carcinoma (n = 61), squamous cell carcinoma (n = 21), adenocarcinoma (n = 40) and small cell lung cancer (n = 11) was 64, 71, 60 and 18%, respectively. Regardless of cell types, the diagnostic sensitivity of CYFRA 21-1 and CEA in malignant pleural effusion (n = 72) was 57 and 60%, respectively (cut-off value of 10 ng/ml in CEA assay). Combining CEA with CYFRA 21-1, the diagnostic sensitivity may increase up to 72%, which was defined at 89% specificity. CONCLUSION: CYFRA 21-1 assay may be a useful tumor marker for discriminating benign from malignant pleural effusion, especially in those of non-small cell lung cancer. The combined use of CEA and CYFRA 21-1 assay in the malignant effusion may increase the diagnostic yield compared with CEA or CYFRA 21-1 alone.     

肿瘤标志物对癌性胸腔积液诊断的价值

目的探讨肿瘤标志物在癌性胸腔积液诊断中的价值。方法应用免疫放射分析法对108例癌性胸腔积液和90例良性胸腔积液的CYFRA21-1、CEA、NSE和CA153含量进行检测,并对结果进行比较分析。结果癌性胸腔积液中四种肿瘤标志物指标明显高于良性组(P<0.01),四项联检后的敏感性、特异性及准确性明显升高。结论肿瘤标志物CYFRA21-1、CEA、NSE和CA153四项联检在癌性胸腔积液的诊断中具有明确的意义。     

Tumor markers in pleural effusion diagnosis

In order to discriminate between malignant and benign effusions, the values of carcinoembryonic antigen (CEA), ferritin, beta2-microglobulin (BMG), acid-soluble glycoprotein (ASP), tissue polypeptide antigen (TPA), adenosine deaminase (ADA), and immunosuppressive acidic protein (IAP) were measured in the pleural fluid of 54 patients with lung cancer, 20 with malignancies other than lung cancer, 18 with tuberculous pleurisy, and 22 with benign diseases other than tuberculosis. CEA levels in malignant effusions were significantly higher than those in benign effusions. At a cutoff level of 5 ng/ml, 68% of the patients with lung cancer and 44% of the patients with other malignancies showed elevated pleural fluid CEA levels. In 13 lung cancer cases with negative pleural fluid cytology, nine cases had elevated pleural fluid CEA levels. The mean pleural fluid BMG level of patients with benign diseases was significantly higher than that of patients with malignant diseases, but there was a marked overlap between those with malignant and benign diseases. No significant differences were found in the pleural fluid ferritin, ASP, TPA, and IAP levels between malignant and benign conditions. ASP and IAP pleural fluid levels showed significant correlations with the pleural fluid C-reactive protein (CRP) concentrations suggesting that they also reflect inflammatory activity. The mean ADA activity in tuberculous effusion was significantly higher than that resulting from other causes of pleural effusion.     

Diagnostic value of CA 549 in pleural fluid. Comparison with CEA,CA 15.3 and CA 72.4

Several tumor markers have been evaluated in pleural fluid, but their clinical role has not been firmly established. The aim of this study is to determine the diagnostic value of carbohydrate antigen 549 (CA 549) levels in pleural fluid, and to compare it with another previously studied tumor markers: carcinoembryonic antigen (CEA), CA 15.3 and CA 72.4. We prospectively studied 252 patients with pleural effusion: 101 malignant (20 mesothelioma) and 151 of several benign diseases. The levels of the tumor markers were measured by immunoradiometric assays (RIA). CA 549 in pleural fluid has an acceptable sensitivity (0.49), with high specificity (0.99). The best combination of tumor markers for differentiating malignant from benign effusions was CA 549+CEA+CA 15.3, with a sensitivity of 0.65, specificity of 0.99 and accuracy of 0.85. The addition of any one tumor marker assay consistently improved the diagnostic value of cytology. In our study, none of the tumor markers was organ-specific. When mesothelioma and hematological malignancy were ruled-out, the combination of CA 549+CEA+CA 15.3, improved the results up to a sensitivity of 0.77, specificity of 1 and accuracy of 0.92. In conclusion, CA 549 assay has an acceptable sensitivity with high specificity. The best combination of tumor markers in this series with a high relative frequency of mesothelioma and low frequency of breast carcinoma was CA 549+CEA+CA 15.3. Individual tumor markers or their combination increased the sensitivity of pleural cytology.     

Clinical evaluation of four tumor markers in malignant and benign pleural effusions.

Total sialic acid (TSA), lipid-bound sialic acid (LSA), ferritin and carcinoembryonic antigen (CEA) were evaluated in 55 patients with malignant pleural effusions and in 32 patients with benign (exudative) pleural effusions. No significant differences were found in the pleural fluid TSA, LSA and ferritin levels between malignant and benign conditions. CEA levels in malignant effusions were significantly higher than those in benign effusions (43.13 +/- 72.8 ng/ml versus 2.6 +/- 5.56 ng/ml, p less than 0.01). At a cut-off level of 5 ng/ml, 60% of the patients with cancer showed elevated pleural fluid CEA levels. The specificity and diagnostic accuracy of CEA in distinguishing malignant from benign pleural exudates were both very high (91% and 71% respectively). Therefore, of the four markers investigated, only CEA could be a valuable tool in the detection of pleural malignancy.     

Diagnostic value of carbohydrate antigens in supernatants and sediments of pleural effusions

A panel of tumour markers including carcinoembryonic antigen (CEA), carbohydrate antigen (Ca)15-3, Ca125 and Ca19-9 were measured in the lysate of sediments and in the supernatants of pleural effusions of patients with benign and malignant disease. The tumour markers were also measured in the serum of the same patients. Of these patients, 32 had benign diseases (12 trasudative effusions associated with cirrhosis and 20 with non-malignant exudates: 12 pleuritis and 8 other inflammations) and 103 had malignant effusions (37 breast cancers, 29 lung cancers, 10 ovary cancers, 6 kidney cancers, 11 mesotheliomas and 10 lymphomas). We showed the highest level of CEA in pleural effusions of lung cancer followed by that in pleural effusions of breast cancer; whereas Ca15-3 was very high in the pleural effusions of breast and lung cancer. Concerning the lysate of sediment, CEA was high in the pleural effusions of patients with lung cancer and Ca15-3 in those of patients with breast cancer. The other markers are much less useful. For the remaining tumours, none of the markers tested appear to aid in the diagnosis of disease. In conclusion, our data suggest that the combined determination of tumour markers on supernatants and sediments of pleural effusion may provide additional information on the nature of pleural effusion, especially for cases with negative cytology.     

Clinical evaluation of the simultaneous determination of tumor markers in fluid and serum and their ratio in the differential diagnosis of serous effusions.

We evaluated the diagnostic utility of simultaneous determination of 5 tumor markers, CEA, CA 125, CA 15-3, CA 19-9 and cytokeratin 19 (CYFRA 21-1), in fluid and serum from 101 patients, 52 with pleural effusion (22 malignant) and 49 patients with ascites (14 malignant). Tumor marker concentrations in fluid from patients with malignant effusions were significantly higher than those obtained in benign fluids or serum. However, there are two types of tumor markers: those released/secreted by normal mesothelia such as CA 125 and cytokeratin 19 (higher levels in benign fluids than in serum) and non-released/secreted tumor markers (low concentrations in benign fluids) such as CEA, CA 19-9 and CA 15-3. The fluid/serum (F/S) ratio showed better sensitivity with maximum specificity than a single determination in fluid for CEA, CA 15-3 and CA 19-9, but not for CA 125 and CYFRA. The combination of a F/S ratio greater than 1.2 and a cut-off of 5 ng/ml for CEA, 30 U/ml for CA 15-3 and 37 U/ml for CA 19-9 showed sensitivities of 58, 57 and 44%, respectively, and a specificity of 100%, with a combined sensitivity of 82% for overall effusions and 79% for fluids with negative cytology with a specificity of 100%. In conclusion, the use of the F/S ratio in nonsecreted tumor markers such as CEA, CA 19-9 and CA 15-3 improve the sensitivity and specificity and allow standardization of the cut-off.     

胸水中CYFRA21-1测定的初步报告

 测定了130例胸水标本，其中癌性购水75例，结核性胸膜炎43例，其它良性购水12例。恶性胸水的CYFRA21-1浓度显著高于良性胸水（P＜0．01），其敏感性为61．3%，特异性为87．3%，准确率为72．3%，提示测定胸水CYFRA21－1对恶性胸水的诊断和鉴别诊断具有临床应用价值。     

Tumor markers in the diagnosis of ascitic and pleural puncture fluids

The identification of carcinoma cells in ascitic and pleural effusions by the detection of tumor markers was studied using an indirect immunoperoxidase method. Punctates from 56 patients with carcinomas, 18 with other malignant diseases and 10 with benign diseases were examined. In 35% of carcinomatous punctates tumor cells were stained by a CEA antiserum. No positive cells were found, however, in non-carcinomatous benign or malignant effusions. The immunocytochemical staining of CEA could detect tumor cells in 36% of carcinoma cases classified suspicious or negative using conventional cytology only. A TPA antiserum as well as the monoclonal antibody OC 125 stained cells in carcinomatous as well as in non-carcinomatous malignant and in benign punctates.     

Diagnostic value of CYFRA 21-1, CEA, CA 19-9, CA 15-3, and CA 125 assays in pleural effusions: analysis of 116 cases and review of the literature

Levels of tumor markers in pleural effusions may help to establish the diagnosis of pleural malignancy, but the precise diagnostic value of each marker remains unclear. The aim of this study was to assess the diagnostic value of five common pleural fluid tumor markers, carcinoembryonic antigen (CEA), cytokeratin fragment (CYFRA) 21-1, cancer antigen (CA) 15-3, CA 19-9, and CA 125, and to review the literature from the past 15 years. Pleural fluid samples were collected prospectively from 116 patients and assayed for CEA, CYFRA 21-1, CA 15-3, CA 19-9, and CA 125 levels. A MEDLINE search of the English-language literature from the past 15 years was also done. Effusions were classified as benign or malignant on the basis of their definitive pathologic or cytologic diagnoses. The levels of all pleural tumor markers were statistically significantly higher in the malignant group than in the benign group. The marker with the highest accuracy was CEA (85.3%); CA 15-3, CYFRA 21-1, and CA 19-9 had similar accuracies (75.2%, 72.4%, and 71.5%, respectively), and CA 125 had the lowest accuracy (40.5%). On univariate analysis, tumor-marker combinations did not result in a greater accuracy than that of CEA alone. On multivariate logistic regression, CA 15-3 and CYFRA 21-1 were significant predictors of malignancy. Among the nine reports in the literature comparing 11 different tumor markers, CEA, CA 15-3, and CYFRA 21-1 yielded the best results. We conclude that pleural fluid analysis should include CEA for the diagnosis of malignancy. CA 15-3 and CYFRA 21-1 may serve as alternative options.     

六种肿瘤标志物在肺癌胸腔积液中的诊断价值

目的通过对胸腔积液和血清中6种肿瘤标志物的检测及胸腔积液脱落细胞学检查,探讨各指标在肺癌胸腔积液中的诊断价值。方法应用化学发光法和酶联免疫分析法测定50例肺癌和30例肺良性疾病患者的胸腔积液和血清中的癌胚抗原（CEA）、糖类抗原19—9（CA19—9）、鳞状细胞癌抗原（SCC）、神经元特异性烯醇化酶（NSE）、细胞角蛋白19片段（CYFRA21—1）、胃泌素前体释放肽（ProGRP）水平,同时对胸腔积液标本进行脱落细胞学检查,并根据受试者工作特性曲线（ROC）建立合理的临床判断临界值。结果肺癌患者胸腔积液中6种肿瘤标志物水平均高于肺良性疾病者,其中CEA、CA19-9、CYFRA21—1、ProGRP水平显著高于肺良性疾病组（P〈0．05）。胸腔积液CEA、血清CYFRA21—1及CEA含量在胸腔积液与血清中的比值（P／S）在各组中的ROC曲线下面积最大。结论胸腔积液CEA、血清CYFRA21—1及CEA的P／S值在鉴别良、恶性胸腔积液中有一定的辅助诊断价值,胸腔积液CEA的诊断价值最大。     

流式细胞术在恶性胸腔积液诊断中的价值

目的 探讨胸腔积液脱落细胞ＤＮＡ定量分析对恶性胸腔积液的诊断价值。方法 采用流式细胞仪（ＦＣＭ）对７２例恶性胸腔积液和５８例良性胸腔积液做ＤＮＡ定量分析,并同进与胸膜活检病理诊断,人积液细胞学诊断及胸腔积液ＣＥＡ测定等方法相比较。结果 以异倍体一为标准,ＦＣＭ诊断恶性胸腔积液的敏感性为８０．６％,特异性为１００．０．％,与胸膜活检病理诊断（８６．１％）。无显著性差异（Ｐ〉０．０５）,但显著高于胸腔     

New approaches in the diagnostic procedure of malignant pleural effusions

The content of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (Ca 19-9), carbohydrate antigen 15-3 (Ca 15-3) and the expression of LewisY related carbohydrate antigens in benign and malignant pleural effusion were determined. These included 35 malignant pleural effusions: 13 breast cancers, 12 lung cancers (6 squamous cell carcinomas, 5 adenocarcinomas and 1 microcytoma), 2 mesotheliomas, 1 epithelioma, 1 kidney cancer, 1 hepatocarcinoma, 1 colon carcinoma, 3 lymphomas, 1 osteosarcoma and 9 benign pleural effusions. We showed that pleural fluid content of CEA, Ca 19-9 and Ca 15-3 were higher in malignant than in benign effusions. However CEA levels in squamous lung cancers were very high in both serum and pleural fluids whereas its levels were only slightly above the cut-off in breast cancers and in lung adenocarcinomas. Serum and pleural fluid Ca 15-3 values were higher in breast and in lung cancers with the highest values in the patients with breast cancer. Furthermore, the LewisY related carbohydrate antigens, evaluated by the reactivity of the cell extracts to MAb B3, were expressed only in breast cancers. These data suggest that pleural fluid content of CEA, and Ca 15-3 associated with the immunoblotting of cell extracts with MAb B3 appear to be very useful to improve the diagnosis of malignant pleural effusions.