The invention relates to the field of 
biomedicine, in particular to an 
enzyme-linked immunization 
diagnostic reagent kit for detecting an 
enterovirus (EV) 71-type 
antibody (immune 
globulin M (IgM)), and a preparation method and application of the 
diagnostic reagent kit. The probability of hand-foot-and-mouth 
disease and severe infection (
viral encephalitis, viral 
cerebrospinal meningitis and 
pulmonary edema) caused by EV71 type is relatively higher, and 
case fatality rate is relatively higher and can be 10 to 25 percent. The 
enzyme-linked immunization 
diagnostic reagent kit of the EV71-
IgM antibody can be used for diagnosing the infection of the EV71 type. According to related documents about the detection of the EV71-IgM, EV71 
virus cultures serving as indirect 
enzyme-linked immuno 
sorbent assay (ELISA) of envelope antigens has defects in such aspects as specificity, sensitivity and stability, and due to high cultivation cost and low efficiency, a large amount of 
virus cannot be supplied to the market. In order to overcome the defects, the invention provides the 
reagent kit which is used for detecting the EV71-IgM in 
human blood serum, required by clinical examination, simple and convenient to operate and applicable to all medical 
disease control departments, and the preparation method and the application of the 
reagent kit. The invention has the technical scheme that: firstly, the 
human blood serum is added into a micro-pore plate, wherein the 
IgM antibody is obtained by an anti-mu chain which is pre-enveloped on the micro-pore plate, and other uncombined components are washed and removed; secondly, an enzyme labeling object is added, the EV71-IgM in the obtained IgM can be combined with the specificity of an EV71 
recombinant antigen which is labeled by 
horse radish peroxidase (HRP), and after washing, the HRP can react with substrates which are added subsequently; and finally, the aim of detecting the EV71-
IgM antibody is fulfilled.