The invention discloses a kit for inspecting tumour M2 type pyruvate kinase and a preparation method of the kit. The kit comprises a test paper card. The test paper card comprises a bottom plate, thebottom plate is provided with a sample pad, a combination pad, a nitrocellulose membrane and an absorbent pad which are sequentially connected at the ends in the sample flowing direction, capture antibodies marked by quantum dots are adsorbed by the combination pad, the nitrocellulose membrane is successively provided with an inspection belt and a quality control belt in the sample flowing direction, the inspection belt is covered with inspection antibodies, and the quality control belt is covered with IgG; the capture antibodies and the inspection antibodies are anti-tumour M2 type pyruvate kinase antibodies, and the capture antibodies and the inspection antibodies are combined with different epitopes of the tumour M2 type pyruvate kinase; and before the capture antibodies are marked by the quantum dots, the capture antibodies are modified with monophosphate sugar. The kit has the advantages that high sensitivity and specificity are achieved, operation is fast, simple and convenient,the result is accurate, and affordability is achieved.