The invention provides amplification primer sets of Klebsiella pneumoniae, application and a method for detecting the Klebsiella pneumoniae. The method for detecting the Klebsiella pneumoniae comprises the following steps: 1) extracting a genome of a Klebsiella pneumoniae sample to be detected; 2) providing the following specific gene rcsA amplification primer sets of the Klebsiella pneumoniae: CP1, CP2, F1, F2, D1, C1, R1, D2, C2, R2, C1*, C2*, D1* and D2*; 3) using genomic nucleic acid of the Klebsiella pneumoniae sample to be detected as a template for isothermal amplification under a multi-crossover isothermal amplification reaction system comprising the amplification primer sets to obtain an amplification product; 4) detecting the amplification product of the step 3) by using a gold nanobiosensor. The method provided by the invention uses multi-crossover isothermal amplification as a basis, is combined with a nanometer biological detection technology, and can rapidly, sensitivelyand highly specifically identify the Klebsiella pneumoniae.