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目的应用流式细胞仪检测胸腔积液中的DNA、RNA、增殖细胞核抗原(PCNA),探讨多参数流式细胞术对恶性胸腔积液的诊断价值。方法2003年8月至2004年2月在我院呼吸内科住院治疗的胸腔积液患者47例,其中19例非肿瘤性胸腔积液患者作为对照组,28例经病理学检查确诊的恶性胸腔积液患者为试验组。采用碘化丙啶染色检测DNA,哌若宁染色检测RNA,PCNA-FITC法检测PCNA,阴性对照采用鼠-α-2a。用美国Becton Dickinson公司FacS Calibur流式细胞仪进行检测,计算单项检测和联合检测的敏感性和特异性。结果(1)非肿瘤性胸腔积液中DNA指数、RNA指数、PCNA流式细胞术检测结果分别为1.03±0.06、10.03±0.54及(4.86±0.72)%,而恶性胸腔积液为1.26±0.17、11.65±1.45及(11.97±1.50)%。诊断分界点分别为1.10%、10.75%、4.56%,此时的敏感性分别为89.3%、78.6%、75.0%,特异性为89.5%、98.5%、84.2%;(2)恶性胸腔积液中DNA指数正常而RNA指数异常者6例,表明流式细胞术同时检测患者RNA可以弥补DNA检测的不足;(3)5例患者胸腔积液中细胞学检查未发现肿瘤细胞,但其DNA指数和RNA指数均高于正常值,经多次胸膜活检或肺部肿块穿刺证实为恶性胸腔积液,表明流式细胞术对细胞学检查有补充作用;(4)DNA指数+RNA指数、DNA指数+PCNA的流式细胞术检测结果、RNA指数+PCNA的流式细胞术检测结果及三者联合诊断的敏感性分别为98.2%、89.3%、89.3%及92.9%,特异性为84.2%、89.5%、84.2%及94.2%。三者联合检测具有较低的漏诊率和误诊率,而对照组未发现三者同时高于诊断临界点的患者。结论流式细胞术检测胸腔积液DNA指数、RNA指数、PCNA的流式细胞术检测结果对于恶性胸腔积液的诊断具有一定的价值,特别是对于部分细胞学检测阴性的恶性胸腔积液的诊断可能具有重要的临床意义。DNA、RNA同时检测对于诊断DNA正常而RNA发生异常改变的恶性胸腔积液具有重要价值,可以弥补单项DNA检测的不足。三者联合检测对于恶性胸腔积液的诊断价值大于单项或两项联合检测,具有较低的漏诊率和误诊率。
Objective To analyze the DNA, RNA and proliferating cell nuclear antigen (PCNA) in pleural effusions by flow cytometry, and to explore the diagnostic value of multiparameter flow cytometry for malignant pleural effusion. Methods Forty-seven cases of pleural effusions hospitalized in our Department of Respiratory Medicine from August 2003 to February 2004 were included. Among them, 19 patients with non-neoplastic pleural effusion served as control group, and 28 patients with malignant pleural cavity diagnosed by pathological examination. The liquid patient was the test group. DNA was detected by propidium iodide staining, RNA was detected by piperazine staining, PCNA was detected by PCNA-FITC method, and mouse-α-2a was used in the negative control. The detection was performed on a FacS Calibur flow cytometer from Becton Dickinson, USA, and the sensitivity and specificity of single detection and combined detection were calculated. Results (1) The results of DNA index, RNA index, and PCNA flow cytometry in non-neoplastic pleural effusions were 1.03±0.06, 10.03±0.54, and (4.86±0.72)%, respectively, while the malignant pleural effusion was 1.26±0.17. , 11.65±1.45 and (11.97±1.50)%. The diagnostic cut-off points were 1.10%, 10.75%, and 4.56%, respectively. The sensitivity at this time was 89.3%, 78.6%, and 75.0%, and the specificity was 89.5%, 98.5%, and 84.2%. (2) Malignant pleural effusion There were 6 cases with normal DNA index and abnormal RNA index, indicating that the simultaneous detection of RNA by flow cytometry could make up for the lack of DNA detection; (3) No cytology was found in cytology of pleural effusion in 5 patients, but their DNA index and The RNA index was higher than normal and was confirmed as a malignant pleural effusion after multiple pleural biopsy or lung puncture. This indicates that flow cytometry complements cytology; (4) DNA index + RNA index, DNA index + The flow cytometry results of PCNA, flow cytometry results of RNA index+PCNA, and the sensitivity of the three combined diagnosis were 98.2%, 89.3%, 89.3%, and 92.9%, respectively, and the specificity was 84.2%, 89.5%. 84.2% and 94.2%. The combined detection of the three had a low missed diagnosis rate and misdiagnosis rate, while the control group did not find any patients whose diagnostic thresholds were higher than the three at the same time. Conclusion Flow cytometric detection of pleural effusion DNA index, RNA index, and PCNA flow cytometry results have certain value in the diagnosis of malignant pleural effusions, especially for the diagnosis of malignant pleural effusions that are negative for some cytological tests. It may have important clinical significance. Simultaneous detection of DNA and RNA is of great value in the diagnosis of malignant pleural effusions with normal DNA and abnormal RNA changes, which can compensate for the inadequacy of individual DNA detection. The combined detection of these three methods was more effective than single or combined detection in the diagnosis of malignant pleural effusion, with a low rate of missed diagnosis and misdiagnosis.