逆转录酶环介导的等温扩增与逆转录酶聚合酶链反应检测前列腺特异性抗原的比较

Mohammad Almasi, M. Esmaili
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引用次数: 0

摘要

背景:研究表明,前列腺癌在男性最常见的五大癌症中排名第二。已经证实,当循环前列腺特异性抗原(PSA)转录物被成功检测到时,可以在早期诊断前列腺癌细胞。建立了一种逆转录酶环介导的等温扩增(RT-LAMP)检测方法,并将其与逆转录酶聚合酶链反应(RT-PCR)检测PSA进行了比较。方法:选取47例患者,其中前列腺癌患者30例,良性前列腺增生(BPH)患者15例,健康对照2例。将2例患者的前列腺癌细胞系(PC3和LNCaP)作为阳性对照纳入研究。然后,从新鲜样品中提取RNA,使用第一链cDNA合成试剂盒进行cDNA合成。采用人前列腺特异性抗原基因设计特异性引物。结果:对照组和BPH患者均无阳性反应。15例局限性癌患者中有13例(86.6%)PSA阳性。所有15例转移患者和阳性对照(100%)均观察到PSA阳性结果。RT-LAMP是一种有利的方法,因为它具有高灵敏度(1000倍),相当便宜,用户友好且安全;此外,可以使用GineFinderTM染料在水浴中快速进行视觉检测。结论:在基础仪器的辅助下,RT-LAMP技术应用简单、可靠,实验结果可直观检验。
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Comparison of Reverse Transcriptase Loop-Mediated Isothermal Amplification and Reverse Transcriptase Polymerase Chain Reaction for Detection of Prostate Specific Antigen
Background: Research shows that prostate cancer ranks second among the top five most common cancers in men. It has been confirmed that when circulating Prostate Specific Antigen (PSA) transcripts are successfully detected, prostate cancer cells can be diagnosed at an early stage. A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was developed and compared to reverse transcriptase polymerase chain reaction (RT-PCR) assay for detection of PSA. Methods: 47 patients, including 30 patients with prostate cancer, 15 with Benign Prostate Hyperplasia (BPH) and 2 healthy subjects as negative controls were included in this study. The prostate cancer cell lines (PC3 and LNCaP) of two patients were included in the study as positive controls. Next, RNA was extracted from fresh samples and a first strand cDNA synthesis kit was applied for the synthesis of cDNA. The human prostate specific antigen gene was used to design specific primers. Results: The results indicated that the control subjects and participants suffering from BPH were not positive. 13 out of 15 (86.6%) patients suffering from localized cancer were PSA positive. PSA positive results were observed among all 15 metastatic patients and positive controls (100%). RT-LAMP is an advantageous method because it is highly sensitive (1000-fold), quite cheap, user-friendly, and safe; in addition, it can be quickly performed by visual detection using GineFinderTM dye in a water bath. Conclusion: RT-LAMP technique can be simply and reliably applied with the aid of basic instruments, and its results can be visually inspected in laboratory studies.
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