This
paper presents a peptide-mediated immunomagnetic separation
technique and an immunofluorescence quantum dot technique for simultaneous
and rapid detection of
Escherichia coli
O157:H7
,
Staphylococcus aureus
, and
Vibrio parahaemolyticus
. First,
three peptides that can specifically recognize the three foodborne
pathogens were combined with magnetic nanoparticles to form an immunomagnetic
nanoparticle probe for capturing three kinds of target bacteria and
then added three quantum dot probes (quantum dots-aptamer), which
formed a sandwich composite structure. When the three quantum dot
probes specifically combined with the three pathogenic bacteria, the
remaining fluorescent signal in the supernatant will be reduced by
magnetic separation. Therefore, the remaining fluorescent signal in
the supernatant can be measured with a fluorescence spectrophotometer
to indirectly determine the three pathogens in the sample. The linear
range of the method was 10–10
7
cfu/mL, and in the
buffer, the detection limits of
E. coli
O157:H7
,
S. aureus
, and
V. parahaemolyticus
were 2.460,
5.407, and 3.770 cfu/mL, respectively. In the tap water simulation,
the detection limits of
E. coli
O157:H7
,
S. aureus
, and
V. parahaemolyticus
were 2.730, 1.990 × 10
1
, and 4.480 cfu/mL, respectively. In the milk simulation sample,
the detection limits of
E. coli
O157:H7
,
S. aureus
, and
V. parahaemolyticus
were 6.660, 1.070 × 10
1
, and 2.236 × 10
1
cfu/mL, respectively. The
method we presented can detect three kinds of foodborne pathogens
at the same time, and the entire experimental process did not exceed
4 h. It has high sensitivity and low detection limit and may be used
in the sample detection of other pathogens.