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首页> 外文期刊>BJU international >Highly specific detection of prostate-specific antigen-positive cells in the blood of patients with prostate cancer or benign prostatic hyperplasia, using a real-time reverse-transcription-polymerase chain reaction method with improved sensitivity.
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Highly specific detection of prostate-specific antigen-positive cells in the blood of patients with prostate cancer or benign prostatic hyperplasia, using a real-time reverse-transcription-polymerase chain reaction method with improved sensitivity.

机译：使用实时逆转录-聚合酶链反应方法提高敏感性，可高度特异性地检测前列腺癌或良性前列腺增生患者血液中的前列腺特异性抗原阳性细胞。

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OBJECTIVE: To assess the presence of circulating prostate-specific antigen (PSA)-expressing cells in patients with prostate cancer or benign prostatic hyperplasia (BPH), and to determine their diagnostic usefulness using a highly sensitive quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) method. PATIENTS, SUBJECTS AND METHODS: Venous blood samples were obtained from 175 patients with prostate cancer (12 metastatic and 163 not metastatic), 49 with BPH, and 50 healthy volunteers. To improve the specificity and sensitivity of the qRT-PCR three innovative features were combined; a primer overlapping two adjacent exons to inhibit nonspecific amplification; a no-end-point first round amplification to increase the sensitivity; and a target-specific primer for the RT phase to increase the specificity. RESULTS: The sensitivity of the method was 1 cell/mL of blood and the interassay coefficient of variation was 10.5%. None of the healthy subjects tested positively, while 9% of those with prostatic cancer and 14% with BPH had PSA-positive cells in the blood. There was a positive association between a positive test and the National Comprehensive Cancer Network classification in the patients with newly diagnosed prostate cancer (P = 0.022). There were no additional statistically significant associations. CONCLUSION: Our results strongly indicate that although there were no false-positive results and the sensitivity of the method was increased to maximal levels, a low frequency of positive results in patients with prostatic cancer and a high frequency of positive results in those with BPH seems to discourage the use of PSA-positive circulating cells in the search for a clinical diagnosis of prostate cancer.
机译：目的：评估前列腺癌或良性前列腺增生（BPH）患者中循环前列腺特异性抗原（PSA）表达细胞的存在，并使用高度灵敏的实时定量逆转录聚合酶链反应来确定其诊断价值反应（qRT-PCR）方法。患者，受试者和方法：静脉血样本来自175例前列腺癌患者（12例转移和163例非转移性），49例BPH和50名健康志愿者。为了提高qRT-PCR的特异性和灵敏度，将三个创新功能进行了组合;重叠两个相邻外显子的引物，以抑制非特异性扩增;无终点的第一轮扩增以提高灵敏度; RT阶段的靶标特异性引物可提高特异性。结果：该方法的灵敏度为1细胞/ mL血液，批间变异系数为10.5％。健康受试者均未检测出阳性，而前列腺癌受试者中有9％，BPH受试者中有14％血液中PSA阳性细胞。在新诊断的前列腺癌患者中，阳性试验与国家综合癌症网络分类之间存在正相关关系（P = 0.022）。没有其他具有统计意义的关联。结论：我们的结果有力地表明，尽管没有假阳性结果，并且该方法的灵敏度提高到最大水平，但前列腺癌患者的阳性结果发生率较低，而前列腺增生患者的阳性结果发生率较高不鼓励使用PSA阳性循环细胞来寻找前列腺癌的临床诊断。

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