The invention discloses a kit and method for detecting Acinetobacter baumannii DNA; the kit comprises: a nucleic acid release agent, a PCR (polymerase chain reaction) liquid, an enzyme mixed liquid, an Acinetobacter baumannii positive control, and an Acinetobacter baumannii negative control, wherein the PCR liquid is made from PCR buffer solution, deoxyribonucleoside, primers for amplifying targeted polynucleotide, a probe for amplifying targeted polynucleotide, primers for amplifying internally labeled fragments and a probe for detecting an internal label. By optimizing conditions in the embodiment of the invention, primers for amplifying targeted polynucleotide and probes for amplifying the targeted polynucleotide, both having optimal amplification, are screened out, the kit is capable of detecting not pathogens of Acinetobacter baumannii but various infections of Acinetobacter baumannii but, is highly specific, and accordingly has greatly improved detection sensitivity, accuracy and stability.