Loop-mediated isothermal amplification (LAMP) of nucleic acids enables the detection of amplification products 10–20 min after the beginning of the reaction. An intraoperative method for the detection of metastases in lymph nodes based on LAMP, one-step nucleic acid amplification (OSNA), allows the rapid detection of the epithelial-specific marker gene KRT19 in lymph nodes. The standard protocol for OSNA was developed more than 10 years ago to detect breast cancer metastases in lymph nodes. Since then, a new version of the key enzyme involved in the reaction (Bst-polymerase) was obtained, but its use in OSNA has remained 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. In this work, the LAMP protocol was developed for the new Bst 3.0 polymerase with optimized dNTP concentrations and without reverse transcriptase, and cell lines were selected (DU-145 for prostate cancer and Molt-4 for negative control). As a result, a new, in vitro system for the LAMP-based detection of prostate cancer with the marker gene KRT19 was developed.

环介导的核酸等温扩增（LAMP）可在反应开始后10–20分钟检测扩增产物。一种基于LAMP的术中检测淋巴结转移的方法，即一步核酸扩增（OSNA），可以快速检测淋巴结中的上皮特异性标记基因KRT19。OSNA的标准协议是10多年前开发的，用于检测淋巴结中的乳腺癌转移。从那时起，新版本的关键酶参与了反应（Bst-聚合酶），但其在OSNA中的用途仍未开发。而且，将OSNA应用于其他癌症类型（尤其是前列腺癌）的检测的时机已经成熟。第一步是创建一个体外系统，该系统可以在前列腺癌细胞系上进行LAMP。在这项工作中，针对具有优化的dNTP浓度且没有逆转录酶的新型Bst 3.0聚合酶开发了LAMP方案，并选择了细胞系（DU-145用于前列腺癌，Molt-4用于阴性对照）。结果，开发了一种新的体外系统，用于使用标记基因KRT19进行基于LAMP的前列腺癌检测。