Magnetically actuated physical impingement for elution of artificial mucous from a swab

abstract

Swabs are used as a collecting device for many biological samples and its complete elution is a desired step for clinical and forensic diagnostics. Swabs are made of cotton, rayon, polyurethane, foam or polyester and come in a spun or flocked-tipped format. They are used to extract biological samples from a patient, which includes saliva, mucous, blood, semen or other body fluids. These body fluids then undergo the process of elution where the collected samples are extracted from the swabs into an elution fluid. Apart from biological samples, the importance of swabbing and elution also becomes more evident in forensics, where the concentration of available cells is very low. One such example is rape kit analysis. Another field of application is the capture and release of bacterial spores from environmental contact surfaces and food surfaces, which also indicate the use of swabs in non-biological areas. The recovery of the biological material from the fibre matrix of the swab has a significant influence on diagnostic sensitivity of any assay. The recovery of micro-organisms from a matrix of swab fibres depends on the nature of the body fluid, the type of the swab fibres and the process of elution. Various methods are used to elute samples from swab, including the use of chemicals to digest the cotton fibres to remove intact cells (~20% recovery), centrifugation (~58% recovery), piezoelectric vibration or pressurized fluid-flow (~60% recovery). These methods are either passive (chemical elution) or provides a gentle tangential shear force through associated flow (centrifugation, piezoelectric and pressurized flow), resulting in a low recovery. The success of all the downstream processes of elution, like lysis, DNA amplification and detection, depends on the number of cells eluted from the swab fibre matrix. Hence, the recovery efficiency is an important parameter for determining the performance of elution, and higher value of the same is desired for most diagnostic assays. This thesis reports a magnetically-actuated physical impingement method for elution and recovery of artificial sputum samples from cotton fibres. A device has been fabricated to induce a rotating magnetic field on smaller magnetic particles in a vial for striking the swab within a confined gap. Elution of samples from the swab using this device was demonstrated using artificial sputum prepared by mixing 2% methyl cellulose in deionised water, loaded with fluorescent-tagged polystyrene beads and Escherichia coli bacteria at various concentrations. The recovery efficiency was found to increase with both rotational speed and elution time, but plateaus after 400 RPM and 120s respectively. At higher concentration of polystyrene beads, a maximum recovery of ~85% was achieved at 5x108 particles/ml sample. With lower concentration (106 particles/ml), the maximum efficiency (~93%) was found to be almost twice of the static condition (46.7%), while using only 620µL of elution volume. Similar trends were found in experiments with artificial sputum loaded with E. coli cells, and the maximum recovery was found to be ~90% at 105 CFU/ml concentration. The robust design and smaller size allows the device to be used in different clinical, forensic and laboratory settings. Also, due to cheaper means of manufacturing and assembly, the vials and smaller magnets can be discarded after every experiment, thereby preventing contamination. The device is most suitable for recovering cells from different body fluids like saliva, mucous, semen or blood, absorbed by the swab fibres. Apart from body fluids samples, swabs holding biological agents from environmental surfaces can also be eluted. A higher recovery at lower concentration facilitates the use of this device where the available analyte concentration is low.