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N-glycolyneuraminic acid (Neu5Gc) is abundant sialic acids in many mammals,which is occurring in the glycoconjugates of most deuterostome animals.Neu5Gc also occur in fresh samples of human tumors and fetuses.However,very little is known about the occurrence and biologic function,due to the limitations of available analytical probe for detection methods.In this study,we first report the development of aptamers specific to Neu5Gc by the in-vitro selection process SELEX.After 15 selecting rounds,cloning,sequencing and ELISA analysis,6 different aptamers were selected that specificity for Neu5Gc.In these 6 aptamers,N8 showed the best IC50 value (126.898 ng mL-1) and had a relatively high affinity constant (Ka=6.68×109 M-1).The aptamers selected in this study will provide a novel analytical probe for the development of Neu5Gc biosensor for detecting of Neu5Gc in tissue and serum of relate tumor patients,as well as animal derived food.And also solved the problem of preparation of antibody in the immunological assays and opened a new way for the detection of Neu5Gc in the prevention of food poisoning and the diagnosis and prevention of tumor diseases.