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@ARTICLE{Nischwitz:908476,
author = {Nischwitz, Volker and Stelmaszyk, Lara and Piel, Sandra and
Tiehm, Andreas},
title = {{C}ascade {F}iltration {W}ith {PCR} {D}etection and
{F}ield-{F}low-{F}ractionation {O}nline {W}ith {ICP}-{MS}
for the {C}haracterization of {DNA} {I}nteraction {W}ith
{S}uspended {P}articulate {M}atter},
journal = {Frontiers in Chemistry},
volume = {10},
issn = {2296-2646},
address = {Lausanne},
publisher = {Frontiers Media},
reportid = {FZJ-2022-02629},
pages = {919442},
year = {2022},
abstract = {The variety of applied antibiotics in animal and human
medicine results in the release, development, and spread of
relevant numbers of antibiotic resistance genes (ARGs) in
the environment. The majority of ARGs are present in
intracellular forms (in bacteria). Neglected aspects are
extracellular variants of ARGs (eARGs) and their fragments,
which have been detected in surface-water samples and
sediments. The stability of eARGs is expected to be low;
however, binding to particulate matter is likely to improve
their stability and also affect their transport and
dissemination behavior. Few studies have investigated DNA
particle interactions, mostly via indirect characterization
of adduct formation in model systems but not in real
environmental matrices. Therefore, our study aims at a novel
approach for direct characterization of desoxyribonucleic
acid (DNA) particle interactions using both cascade
filtration and field-flow fractionation. Cascade filtration
with quantitative polymerase chain reaction (qPCR) detection
indicated retention of ARGs on filters with much larger pore
sizes supporting the hypothesis of ARG-particle
interactions. However, artifacts from membrane clogging or
DNA–membrane interaction cannot be excluded. Consequently,
asymmetric flow field-flow fractionation was investigated as
an alternative separation technique with the advantage of
particle separation in a thin channel, reducing the risk of
artifacts. The key method parameters, membrane composition,
molecular weight cut off, and carrier composition, were
systematically investigated using a calf-thymus DNA-spiked
surface-water sample as a model. The results clearly showed
a shift in the elution time of clay particles suggesting the
presence of DNA–clay adducts. Multi-element detection by
inductively coupled plasma mass spectrometry (ICP-MS)
enabled monitoring of clay via the Al, Fe, and Si signals
and DNA via the P signal. Matching peak profiles for the new
fraction in the fractograms of the ARG and DNA-spiked water
sample support adduct formation. Further evidence was
provided by a novel post-channel filtration approach for the
separation of free DNA from DNA–clay adducts.},
cin = {ZEA-3},
ddc = {540},
cid = {I:(DE-Juel1)ZEA-3-20090406},
pnm = {2151 - Terrestrial ecosystems of the future (POF4-215)},
pid = {G:(DE-HGF)POF4-2151},
typ = {PUB:(DE-HGF)16},
pubmed = {35836676},
UT = {WOS:000824557100001},
doi = {10.3389/fchem.2022.919442},
url = {https://juser.fz-juelich.de/record/908476},
}