Evolution of the Probe-Based Loop-Mediated Isothermal Amplification (LAMP) Assays in Pathogen Detection

Abstract

Loop-mediated isothermal amplification (LAMP), as the rank one alternative to a polymerase chain reaction (PCR), has been widely applied in point-of-care testing (POCT) due to its rapid, simple, and cost-effective characteristics. However, it is difficult to achieve real-time monitoring and multiplex detection with the traditional LAMP method. In addition, these approaches that use turbidimetry, sequence-independent intercalating dyes, or pH-sensitive indicators to indirectly reflect amplification can result in false-positive results if non-specific amplification occurs. To fulfill the needs of specific target detection and one-pot multiplex detection, a variety of probe-based LAMP assays have been developed. This review focuses on the principles of these assays, summarizes their applications in pathogen detection, and discusses their features and advantages over the traditional LAMP methods.

Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) for the diagnosis of bacterial periprosthetic joint infection

“…Therefore, we inferred that, overall, the analytical sensitivity of our assay was higher than that reported in previous studies [ 40 ]. The sensitivity to each target does not deteriorate when LAMP is developed as a multiplex assay, as with real-time PCR [ 40 , 41 ]. In our study, the analytical sensitivity was approximately 10–10 2 times lower when each type of bacterium was individually tested in a single-plex assay than in a multiplex assay.…”

DNA Polymerase I Large Fragment from Deinococcus radiodurans, a Candidate for a Cutting-Edge Room-Temperature LAMP

“…To overcome the limitations of PCR in the 1990s, new amplification techniques were developed [29], which allow the synthesis of nucleic acids at a constant temperature, without the need of thermal cyclers. This new technology, known as isothermal amplification, has had extensive development, giving rise to multiple amplification techniques; among these, the most promising is the loop-mediated isothermal amplification (LAMP) [30,31]. This technique involves the use of Bst PolI, a DNA polymerase that, in addition to having synthetic activity, also has strand-displacement activity, indispensable for eliminating the DNA re-opening phase used in PCR during the amplification process.…”

Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of Hermetia illucens (Diptera: Stratiomyidae)

“…This method is particularly suited to developing countries with limited access to expensive testing equipment. Therefore, this approach has emerged as a viable alternative to PCR-based technologies not only in agricultural industry testing but also in other diagnostic assay application fields (the LAMP approach has been successfully utilized for the quick and selective detection of several types of infections) [31][32][33][34][35][36][37][38][41][42][43][44][45][46][47][48][49].…”

“…LAMP is one of the most deeply researched and well-established molecular detection approaches currently available, which provided critical support during the research and development process [34][35][36][37][38][39][40]. In comparison with the PCR-based method, the LAMPbased technique is quite easy; a heating block or water bath should suffice, and the reaction should be carried out at a steady temperature (60-65 • C).…”