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

  • Mohammad Amin Almasi Young Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran
  • Marya Esmaili MSc. Student, Microbiology, University of Zanjan and Expert Laboratory Research Unit of Keshtodam-Dashband Co., Boukan, Iran
Keywords: Benign prostate hyperplasia, Prostate cancer, Prostate specific antigen, RT-LAMP assay, RT-PCR assay


Background: 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.  


Seiden MV, Kantoff PW, Krithivas K, Propert K, Bryant M, Haltom E, Gaynes L, Kaplan I, Bubley G, DeWolf W, Sklar J. Detection of circulating tumor cells in men with localized prostate cancer. Journal of Clinical Oncology. 1994 Dec 1;12(12):2634-9.

Boring CC, Squires TS, Tong T. Cancer statistics, 1994. CA: a cancer journal for clinicians. 1994;44(1):7-26.

Wingo PA, Landis S, Ries LAG. An adjustment to the estimate for new prostate cancer cases. CA A cancer J Clin. 1997;80:1810-13.

Daher R, Beaini M. Prostate-specific antigen and new related markers for prostate cancer. Clinical chemistry and laboratory medicine. 1998 Sep 1;36(9):671-81.

Landis SH, Murray T, Bolden S, Wingo PA. Cancer statistics, 1999. CA: A cancer Journal for Clinicians. 1999 Jan;49(1):8-31.

Gao CL, Rawal SK, Sun L, Ali A, Connelly RR, Banez LL, Sesterhenn IA, Mcleod DG, Moul JW, Srivastava S. Diagnostic potential of prostate-specific antigen expressing epithelial cells in blood of prostate cancer patients. Clinical cancer research. 2003 Jul 1;9(7):2545-50.

Gewanter RM, Katz AE, Olsson CA, Benson MC, Singh A, Schiff PB, Ennis RD. RT-PCR for PSA as a prognostic factor for patients with clinically localized prostate cancer treated with radiotherapy. Urology. 2003 May 1;61(5):967-71.

Lundwall إ, Lilja H. Molecular cloning of human prostate specific antigen cDNA. FEBS letters. 1987 Apr 20;214(2):317-22.

Shariat SF, Kattan MW, Song W, Bernard D, Gottenger E, Wheeler TM, Slawin KM. Early postoperative peripheral blood reverse transcription PCR assay for prostate-specific antigen is associated with prostate cancer progression in patients undergoing radical prostatectomy. Cancer research. 2003 Sep 15;63(18):5874-8.

Wang TJ, Rittenhouse HG, Wolfert RL, Lynne CM, Brackett NL. PSA concentrations in seminal plasma. Clinical chemistry. 1998 Apr 1;44(4):895a-6.

Lilja H. A kallikrein-like serine protease in prostatic fluid cleaves the predominant seminal vesicle protein. The Journal of clinical investigation. 1985 Nov 1;76(5):1899-903.

Stamey TA, Yang N, Hay AR, McNeal JE, Freiha FS, Redwine E. Prostate-specific antigen as a serum marker for adenocarcinoma of the prostate. New England Journal of Medicine. 1987 Oct 8;317(15):909-16.

Brawer MK, Peehl DM, Stamey TA, Bostwick DG. Keratin immunoreactivity in the benign and neoplastic human prostate. Cancer research. 1985 Aug 1;45(8):3663-7.

Israeli RS, Miller WH, Su SL, Powell CT, Fair WR, Samadi DS, Huryk RF, DeBlasio A, Edwards ET, Wise GJ, Heston WD. Sensitive nested reverse transcription polymerase chain reaction detection of circulating prostatic tumor cells: comparison of prostate-specific membrane antigen and prostate-specific antigen-based assays. Cancer research. 1994 Dec 15;54(24):6306-10.

Moreno JG, Croce CM, Rischer R, Monne M, Vihko P, Mulholland SG, Gomella LG. Detection of hematogenous micrometastasis in patients with prostate cancer. Cancer Research. 1992 Nov 1;52(21):6110-2.

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic acids research. 2000 Jun 15;28(12):e63-.

Mori Y, Kitao M, Tomita N, Notomi T. Real-time turbidimetry of LAMP reaction for quantifying template DNA. Journal of biochemical and biophysical methods. 2004 May 31;59(2):145-57.

Tsai SM, Chan KW, Hsu WL, Chang TJ, Wong ML, Wang CY. Development of a loop-mediated isothermal amplification for rapid detection of orf virus. Journal of Virological Methods. 2009 May 1;157(2):200-4.

Almasi MA, Erfan Manesh M, Jafary H, Dehabadi SM. Visual detection of Potato leafroll virus by one-step reverse transcription loop-mediated isothermal amplification of DNA with the GenefinderTM dye. J Virol Methods. 2003;192:51-4.

Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical biochemistry. 1987 Apr 1;162(1):156-9.

Barry MJ. Prostate-specific–antigen testing for early diagnosis of prostate cancer. New England Journal of Medicine. 2001 May 3;344(18):1373-7.

Garnick MB, Fair WR. Combating prostate cancer. Scientific American. 1998 Dec 1;279(6):74-83.

MOUL JW. Prostate specific antigen only progression of prostate cancer. The Journal of urology. 2000 Jun;163(6):1632-42.

Brandt B, Junker R, Griwatz C, Heidl S, Brinkmann O, Semjonow A, Assmann G, Zنnker KS. Isolation of prostate-derived single cells and cell clusters from human peripheral blood. Cancer research. 1996 Oct 15;56(20):4556-61.

Wang ZP, Eisenberger MA, Carducci MA, Partin AW, Scher HI, Ts’o PO. Identification and characterization of circulating prostate carcinoma cells. Cancer. 2000 Jun 15;88(12):2787-95.

Moreno JG, O’Hara SM, Gross S, Doyle G, Fritsche H, Gomella LG, Terstappen LW. Changes in circulating carcinoma cells in patients with metastatic prostate cancer correlate with disease status. Urology. 2001 Sep 1;58(3):386-92.

Racila E, Euhus D, Weiss AJ, Rao C, McConnell J, Terstappen LW, Uhr JW. Detection and characterization of carcinoma cells in the blood. Proceedings of the National Academy of Sciences. 1998 Apr 14;95(8):4589-94.

Sabile A, Louha M, Bonte E, Poussin K, Vona G, Mejean A, Chretien Y, Bougas L, Lacour B, Capron F, Roseto A. Efficiency of Ber-EP4 antibody for isolating circulating epithelial tumor cells before RT-PCR detection. American journal of clinical pathology. 1999 Aug 1;112(2):171-8.

Muscatelli B, Colombel M, Jouault H, Amsellem S, Mazeman E, Abbou CC, Chopin D. In vitro detection of prostate cancer circulating cells by immunocytochemistry, flow cytometry and RT-PCR PSA. Progres en urologie: journal de l’Association francaise d’urologie et de la Societe francaise d’urologie. 1998 Dec;8(6):1058-64.

Goessl C, Mـller M, Heicappell R, Krause H, Miller K. DNA-based detection of prostate cancer in blood, urine, and ejaculates. Annals of the New york Academy of sciences. 2001 Sep;945(1):51-8.

How to Cite
Almasi MA, Esmaili M. Comparison of Reverse Transcriptase Loop-Mediated Isothermal Amplification and Reverse Transcriptase Polymerase Chain Reaction for Detection of Prostate Specific Antigen. Basic Clin Cancer Res. 11(1):35-44.
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