Detection of microorganisms using recombinase polymerase amplification with lateral flow dipsticks

The demand for point of care testing (POCT) in diagnostics has been increasing, especially in developing countries with low resources and poor settings. Isothermal amplification techniques overcome the need for temperature control, exhibiting advantages beyond polymerase chain reaction related techniques. Recombinase polymerase amplification (RPA) is a sensitive, specific, rapid and simple method for amplification of both DNA and RNA, which has been rapidly developed and widely used in many microorganism detection applications. Combined with lateral flow assay (LFA) for readout, RPA-LFA can be potentially developed as a powerful in-field diagnostic tool for POCT. Here, we review recent achievements of RPA-LFA for the detection of microorganisms, including viruses, bacteria and parasites. A detailed protocol with related materials and reagents for RPA-LFA development is also presented. Future trends of RPA further development, including overcoming current drawbacks and the combination with CRISPR associated techniques are discussed at the end.