The invention discloses a method for qualitative and quantitative testing of intestinal microorganisms and belongs to the technical field of biology. The method comprises the steps that a target microorganism class group, target microorganisms and non-target organisms in a sample to be tested and reference microorganisms not existing in the sample to be tested are determined; a feature area for the target microorganism class group and the target microorganisms is designed; multiple amplification primers of the feature area are designed; after the reference microorganisms and exogenous nucleic acid are added into the sample to be tested, nucleic acid of the microorganisms in the sample to be tested is extracted; the designed multiple primers are utilized for amplifying the nucleic acid of the microorganisms, and a feature sequencing segment is obtained after amplification; the feature sequencing segment is utilized for qualitatively and quantitatively analyzing the microorganisms in the sample to be tested. According to the method, there is no need to perform preculture or proliferation on the microorganisms, multiple known microorganisms in the sample to be tested can be tested at one time at high flux, accuracy and resolution, the testing process is simple and quick, and the procedures are standard.