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Sub-femtomole detection of 16s rRNA from Legionella pneumophila using surface plasmon resonance imaging.
[legionellosis]
Legionellosis
has
been
and
continues
to
be
a
life-threatening
disease
worldwide
,
even
in
developed
countries
.
Given
the
severity
and
unpredictability
of
Legionellosis
outbreaks
,
developing
a
rapid
,
highly
specific
,
and
sensitive
detection
method
is
thus
of
great
pertinence
.
In
this
paper
,
we
demonstrate
that
sub-femtomole
levels
of
16
s
rRNA
from
pathogenic
Legionella
pneumophila
can
be
timely
and
effectively
detected
using
an
appropriate
designed
capture
,
detector
probes
,
and
a
QD
SPRi
signal
amplification
strategy
.
To
achieve
specific
and
sensitive
detection
,
optimal
hybridization
conditions
and
parameters
were
implemented
.
Among
these
parameters
,
fragmentation
of
the
16
s
rRNA
and
further
signal
amplification
by
QDs
were
found
to
be
the
main
parameters
contributing
to
signal
enhancement
.
The
appropriate
design
of
the
detector
probes
also
increased
the
sensitivity
of
the
detection
system
,
mainly
due
to
secondary
structure
of
16
s
rRNA
.
The
use
of
16
s
rRNA
from
L
.
pneumophila
allowed
for
the
detection
of
metabolically
active
pathogens
with
high
sensitivity
.
Detection
of
16
s
rRNA
in
solutions
as
diluted
as
1
pM
at
450
μL
(
0
.
45
femtomole
)
was
achieved
in
less
than
3
h
,
making
our
approach
suitable
for
the
direct
,
timely
,
and
effective
detection
of
L
.
pneumophila
within
man-made
water
systems
.
Diseases
Validation
Diseases presenting
"metabolically active pathogens"
symptom
legionellosis
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