ASSAY USING MULTI-LAYER MEMBRANE TO DETECT MICROBIOLOGICAL TARGET AND METHOD OF MANUFACTURING MULTI-LAYER MEMBRANE

摘要

A membrane, method and system are disclosed for rapid, sensitive and precise detection of an agent suspected of being present in a sample. The agent may be a cell or microorganism, e.g., a single pathogenic bacteria, and the sample may be small, e.g., milliliters of unprocessed environmental water. The sample is processed by filtering it through an asymmetric membrane having multiple layers. One layer has microchannels for capturing the agent and another layer has nanochannels for passing particles smaller than the agent. Amplification reagents, such as loop-mediated isothermal amplification (LAMP) reagents, are load onto membrane so that the microchannels act as nanoreactors, creating quantifiable amplicons within the pores on the exposed surface of the membrane in response to captured agent. The amplicons may be imaged and counted using a fluorescent camera. The membrane is capable of agent capture, concentration, purification, partition, lysis and digital LAMP without off-membrane sample treatments.