The invention relates to a double-
antibody sandwich method for detecting
enterobacter sakazakii in food. The method is characterized in that the
enterobacter sakazakii (ATCC29544) and
lipopolysaccharide (LPS) extracted by
coupling the
enterobacter sakazakii (ATCC29544) with BSA (
bovine serum albumin) are respectively used as
immunogen to simultaneously immunize BALB / c mice, and 11 enterobacter sakazakii
monoclonal antibodies are obtained through immunization, fusion and screening to respectively
label HRP (
horse radish peroxidase) and are paired through the enterobacter sakazakii. A No.7
monoclonal antibody CGMCC No. 7207 is used as a
coating antibody, a No.8
monoclonal antibody CGMCCNO.7208 is used as an
enzyme labeled antibody, a sandwich
ELISA method of the enterobacter sakazakii is established by adopting the ATCC29544 as a
standard product, and LOD is 53000CFU / Ml. By adopting the
monoclonal antibody with high physicochemical property, good uniformity, good specificity and capability of being in
mass preparation, the established sandwich method is high in sensitivity and low in cost, no cross reaction with
salmonella enteritidis, E.coli, E.coliO157: H7,
salmonella typhimurium,
staphylococcus aureus and
listeria monocytogenes exists, and thus a rapid and high efficient analysis means is provided for detecting the enterobacter sakazakii in the food.