Multiplex detection of bacteria on an integrated centrifugal disk using bead-beating lysis and loop-mediated amplification

Abstract

Abstract Although culture-based identification of bacteria is the gold-standard for the diagnosis of infectious diseases, it is time consuming. Recent advances in molecular diagnostics and microfluidic technologies have opened up new avenues for rapid detection of bacteria. Here, we describe a centrifugal-microfluidic chip for the detection of bacteria by integrating the cell lysis, clarification, and loop-mediated amplification (LAMP). The major advantages of this chip are as follows. Firstly, bacteria lysis was innovatively achieved by rotating a pair of magnets to generate bead-beating while the chip was kept stationary during lysis, which simplified the chip design because no additional valve was needed. Secondly, the on-chip assay time was short (within 70 min), which was competitive in emergency situations. Thirdly, results of the analysis can be interpreted by using a fluorescence detector or by the naked-eye, making it versatile in many areas, especially the resource-limited areas. The on-chip limits of detection of six types of bacteria were valued by gel electrophoresis, showing the similar results compared to the bench-top LAMP protocol. This chip can be used for rapid, sensitive, accurate and automated detection of bacteria, offering a promising alternative for simplifying the molecular diagnostics of infectious diseases.