Hand, foot, and mouth disease is an acute infectious disease caused by enterovirus 71 (EV71) or coxsackievirus A16 (CVA16) infection in category C. Most severe cases are caused by enterovirus type 71 (EV71) infection. The clinical manifestations of HFMD are fever with rash on hands, feet, mouth and buttocks in common cases, and some cases may be feverless; severe cases mostly occur in preschool children, and the condition is dangerous with high mortality rate. Some infants and children may suffer from encephalitis, encephalomyelitis, brainstem encephalitis, resulting in cerebral edema, increased intracranial pressure, and life-threatening neurogenic pulmonary edema and circulatory failure in a short period of time. the proportion of patients with severe disease and death rate caused by EV71 is significantly higher than other enteroviruses. The pathogenic diagnosis of HFMD is mainly based on viral nucleic acid and virus-specific antibodies. Molecular biology methods such as RT-PCR viral nucleic acid detection are the direct basis for viral infection, and virus-specific antibodies are the indirect basis, among which enterovirus 71 and coxsackievirus CVA16 IgM antibodies are mainly used as an aid to diagnosis in the acute phase of infection. IgM antibodies are used to detect IgM antibodies produced in human serum, plasma or finger whole blood samples during the acute phase of infection, but cannot be used as the only basis for clinical diagnosis, and patients with negative IgM antibodies should also be tested repeatedly during the onset of infection in conjunction with clinical practice. A 4-fold or more increase in enterovirus type-specific neutralizing antibodies compared to the acute phase indirectly indicates viral infection. Enteroviruses sometimes cause asymptomatic infections, so the actual significance of the experimental results should also be interpreted in the context of the clinical course and epidemiological data.