Comparison of Reverse Transcriptase Loop-Mediated Isothermal Amplification and Reverse Transcriptase Polymerase Chain Reaction for Detection of Prostate Specific Antigen
Comparison of RT-LAMP and RT-PCR for Detection of Prostate Specific Antigen
AbstractBackground: Research shows that prostate cancer ranks second among the top five most common cancers in men. It has been confirmed that if the circulating Prostate Specific Antigen (PSA) transcripts are detected successfully, the prostate cancer cells will be diagnosed early. 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, 15 with Benign Prostate Hyperplasia (BPH) and 2 healthy subjects as negative controls were included in this study. Also, the prostate cancer cell lines (PC3 and LNCaP) of two patients were included in the study as positive controls. Next, using the fresh samples, RNA was extracted and a first strand cDNA synthesis kit was applied for synthesis of cDNA. The human prostate specific antigen gene was used to design specific primers. Results: The results indicated that the control subjects and BPH were not positive. 13 out of 15 (86.6%) patients suffering from were localized cancer 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 is performed quickly by visual detection using GineFinderTM dye in a water bath. Conclusions: RT-LAMP technique can be simply and reliably applied with basic instruments through visual inspection in laboratory studies.
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