Performing PCR and other gene-based assays with this sampler.
It seems reasonable to propose that the sampler could be used to collect airborne particles (to a lower limit of ~2 microns) and detect their genomic material by PCR, ITS rRNA or other gene-based sequencing methods.
While we have not performed PCR on samples collected with this sampler, we have used the same electret to collect (by filtration of exhaled breath) and detect exhaled viruses by PCR; see here for assay and here for extraction.
Electret is also becoming widely used for air sampling of fungi by air filtration, due to its low pressure drop and high collection efficiency. Both our PLoS papers used it to detect allergen, the 2013 as a filter, and the 2016 as an impaction medium.
There are a very large number of considerations in sampling and detecting bioaerosols and all applications require optimisation for specific purposes. We cannot hope to cover this topic. Here, is one recent review, and there are many others.
One thing to note from our experience with electrets is that they differ in many obvious ways such as physical appearance but they also differ in hydrophobicity; that means it can be difficult to get extraction buffers to penetrate. We found the best way to manage this was to put the electret into a tube, put the buffer on top and then centrifuge the buffer into the matrix of the electret. We build special holders for the electret*, so that the buffer could subsequently be spun out of the matrix.
Additionally, electrets can be penetrated by solvents, eg 20-50% ethanol, depending on the electret, so the electret can be penetrated by extraction buffers containing these. Such alcohol / solvent containing buffers also reverse the binding of particles to the fibres, so bound particles themselves can be obtained by wetting the electret in this way and then spinning to separate the electret from the buffer and particles which pellet in the bottom of the tube.
(* these are small sleeves with a porous screen that fit into the top portion of an Eppindorf tube).
As noted under 'electret electrostatic cloth' elsewhere, we have a large amount of electret (we were required to purchase a 1000M roll) and are happy to supply at the cost of postage.
While we have not performed PCR on samples collected with this sampler, we have used the same electret to collect (by filtration of exhaled breath) and detect exhaled viruses by PCR; see here for assay and here for extraction.
Electret is also becoming widely used for air sampling of fungi by air filtration, due to its low pressure drop and high collection efficiency. Both our PLoS papers used it to detect allergen, the 2013 as a filter, and the 2016 as an impaction medium.
There are a very large number of considerations in sampling and detecting bioaerosols and all applications require optimisation for specific purposes. We cannot hope to cover this topic. Here, is one recent review, and there are many others.
One thing to note from our experience with electrets is that they differ in many obvious ways such as physical appearance but they also differ in hydrophobicity; that means it can be difficult to get extraction buffers to penetrate. We found the best way to manage this was to put the electret into a tube, put the buffer on top and then centrifuge the buffer into the matrix of the electret. We build special holders for the electret*, so that the buffer could subsequently be spun out of the matrix.
Additionally, electrets can be penetrated by solvents, eg 20-50% ethanol, depending on the electret, so the electret can be penetrated by extraction buffers containing these. Such alcohol / solvent containing buffers also reverse the binding of particles to the fibres, so bound particles themselves can be obtained by wetting the electret in this way and then spinning to separate the electret from the buffer and particles which pellet in the bottom of the tube.
(* these are small sleeves with a porous screen that fit into the top portion of an Eppindorf tube).
As noted under 'electret electrostatic cloth' elsewhere, we have a large amount of electret (we were required to purchase a 1000M roll) and are happy to supply at the cost of postage.
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