Loop-mediated isothermal amplification (LAMP): real-time methods for the detection of the survivin gene in cancer cells

Abstract

It is well known that the survivin gene is an important inhibitor of apoptosis protein because of its unique diagnostic and prognostic potential discovered in recent years. Considering the high morbidity and metastasis of tumor malignancy, early and accurate detection of the survivin gene plays a pivotal role in anticancer therapies. The loop-mediated isothermal amplification (LAMP) assay strategy was demonstrated by using optimized real-time fluorescence measurements in a single tube at a constant temperature of 62 °C for 30–60 min. The survivin gene was the target sequence amplified by LAMP. By testing serial tenfold dilutions of target DNA ranging from 1 × 10⁶ copies to 1 copy per tube, the developed method showed that the detection limit (analytical sensitivity) for the survivin gene was 10 copies per tube. Our assay strategy is able to offer high amplification efficiency, excellent reproducibility, and cost-effective and simplified operations. Without the requirements for sophisticated equipment and technical skills of the PCR, the LAMP method can add a great deal of value to small-scale hospitals, point-of-care testing, and clinical laboratories.