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Updated: Aug 26, 2026
Sequencing Small Non-coding RNA from Formalin-fixed Tissues and Serum-derived Exosomes from Castration-resistant Prostate Cancer Patients
Published on: November 19, 2019
[Conditions for the development of a program of reverse transcriptase-polymerase chain reaction for molecular staging
L Llanes González1, A Ferruelo Alonso, A Páez Borda
1Unidad de Investigación Urológica, Servicio de Urología, Hospital Universitario de Getafe, Madrid.
Objective:
To determine accurately the conditions for the development of tests using RT-PCR for PSA in patients with prostate cancer.
Method:
Gen amplification of PSA in cultures of the human prostate tumoral cell line LNCaP with RT-PCR under hot-start conditions, and verification through enzyme restriction digestion of the PCR product. Also, calculation of the PCR test limit of detection through serial dilutions of LNCaP cells in peripheral blood mononucleate cells.
Results:
A highly specific, easy to perform, reproducible RT-PCR protocol has been developed. The lowest limit of detection reached was 1 PSA synthesising cell per 10(6) peripheral blood mononucleate cells.
Conclusions:
RT-PCR is a highly sensitive technique that allows detection of small numbers of PSA producing cells in peripheral blood. This experience allows to establish with precision the conditions for the development of RT-PCR tests for PSA.
