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The application provides a Bst DNA polymerase domain, a mutant and application thereof, and belongs to the technical field of molecular biology. The amino acid sequence of the Bst DNA polymerase domain is shown in SEQ ID NO:1. The Bst DNA polymerase mutant is obtained by replacing the natural DNA binding domain of the wild-type Bst DNA polymerase with the amino acid sequence shown in SEQ ID NO:1. The Bst DNA polymerase mutant exhibits extremely high catalytic efficiency on dUTP, and also has excellent hot-start characteristics, and can reduce the false positive rate of LAMP technology to close to zero. The Bst DNA polymerase mutant also has extremely strong complex sample tolerance, and the detection sensitivity of the pathogen is 20 times higher than that of the wild-type Bst enzyme, and can meet the early and accurate diagnosis requirements of trace infection.