The invention discloses a real-time fluorescent quantitative PCR (
polymerase chain reaction) detection method as well as a standard substance and a detection kit for real-time fluorescent quantitative PCR detection. The special standard substance is used for drawing standard curves, real-time fluorescent quantitative PCR detection is performed on an HER2
gene of a to-be-detected sample and a
reference gene, and the detection method is used for clinical
targeted therapy of
breast cancer and
stomach cancer. The invention further provides a
plasmid vector containing a
reference gene segment and a to-be-detected
target gene segment in an equal proportion and taken as the detection standard substance. The invention also provides the real-time fluorescent quantitative PCR kit for detecting the amplification state of the HER2
gene. The kit comprises a PCR
reaction system for detecting the amplification state of the HER2
gene, the standard substance and negative and positive controls. The detection method and the detection kit have the advantages of being convenient to use, high in detection efficiency, cost-saving and the like. Great significance in diagnosis of HER2 positive
breast cancer as well as research and development,
quality control and clinical application of
breast cancer resistant drugs is realized.