Система in vitro для детекции маркеров рака предстательной железы методом LAMP

Abstract

Loop-mediated isothermal amplification (LAMP) of nucleic acids enables detection of amplification products in 10-20 min after the reaction begins. An intraoperative method for detection of metastases in lymph nodes is based on LAMP and is called one-step nucleic acid amplification (OSNA). It allows rapid detection of the epithelial-specific marker KRT19 in lymph nodes. The standard protocol for OSNA was developed more than 10 years ago. Since that, a new version of the key enzyme involved in the reaction (Bst-polymerase) was obtained, but its use in OSNA remains unexplored. Moreover, the time has come to apply OSNA to the detection of other cancer types, in particular prostate cancer. The first step is to create an in vitro system that allows LAMP to be carried out on prostate cancer cell lines. Here, LAMP protocol for the new Bst 3.0 polymerase with optimized dNTPs concentration and without reverse transcriptase was developed, and cell lines (DU-145 for prostate cancer and Molt-4 for negative control) were selected. In summary, a new in vitro system for LAMP-based detection of prostate cancer marker KRT19 was developed. LAMP, OSNA, prostate cancer This work was supported by a grant from the President of the Russian Federation for state support of young Russian scientists (MK-1906.2019.4).