The invention relates to the field of SARS-CoV-2 nucleic acid detection, in particular to a PCR (Polymerase Chain Reaction) target sequence, primers and a probe for detecting infectious SARS-CoV-2 and application of the PCR target sequence, the primers and the probe. The PCR target sequence, the primers and the probe for detecting the infectious SARS-CoV-2, which are provided by the invention, can be used for well distinguishing the infectivity of the SARS-CoV-2, and meanwhile, the PCR target sequence, the primers and the probe have the characteristics of high sensitivity and good repeatability. Compared with a conventional infectious SARS-CoV-2 detection test, the the PCR target sequence, the primers and the probe have the advantages that the test period of infectious SARS-CoV-2 detection is greatly shortened, the requirements on scientific research conditions and equipment are reduced, nucleic acid of virus inactivation caused by virus capsid protein damage and infectious virions are effectively distinguished, the detection accuracy is improved, and the false positive phenomenon occurring in actual detection is reduced.