%0 Journal Article
%A Shlyapnikov, Yuri M.
%A Shlyapnikova, Elena A.
%A Simonova, Maria A.
%A Shepelyakovskaya, Anna O.
%A Brovko, Fedor A.
%A Komaleva, Ravilya L.
%A Grishin, Eugene V.
%A Morozov, Victor N.
%D 2012
%T Rapid Simultaneous Ultrasensitive
Immunodetection
of Five Bacterial Toxins
%U https://acs.figshare.com/articles/journal_contribution/Rapid_Simultaneous_Ultrasensitive_Immunodetection_of_Five_Bacterial_Toxins/2508616
%R 10.1021/ac300567f.s001
%2 https://ndownloader.figshare.com/files/4151545
%K food samples
%K application
%K assay
%K Bacterial ToxinsRapid ultrasensitive detection
%K PCR
%K polymerase chain reaction
%K shock syndrome toxin
%K LOD
%X Rapid ultrasensitive detection of gastrointestinal pathogens
presents
a great interest for medical diagnostics and epidemiologic services.
Though conventional immunochemical and polymerase chain reaction (PCR)-based
methods are sensitive enough for many applications, they usually require
several hours for assay, whereas as sensitive but more rapid methods
are needed in many practical cases. Here, we report a new microarray-based
analytical technique for simultaneous detection of five bacterial
toxins: the cholera toxin, the E. coli heat-labile
toxin, and three S. aureus toxins (the enterotoxins
A and B and the toxic shock syndrome toxin). The assay involves three
major steps: electrophoretic collection of toxins on an antibody microarray,
labeling of captured antigens with secondary biotinylated antibodies,
and detection of biotin labels by scanning the microarray surface
with streptavidin-coated magnetic beads in a shear-flow. All the stages
are performed in a single flow cell allowing application of electric
and magnetic fields as well as optical detection of microarray-bound
beads. Replacement of diffusion with a forced transport at all the
recognition steps allows one to dramatically decrease both the limit
of detection (LOD) and the assay time. We demonstrate here that application
of this “active” assay technique to the detection of
bacterial toxins in water samples from natural sources and in food
samples (milk and meat extracts) allowed one to perform the assay
in less than 10 min and to decrease the LOD to 0.1–1 pg/mL
for water and to 1 pg/mL for food samples.
%I ACS Publications