Helicase-dependent Isothermal DNA Amplification (HDA) is an isothermal amplification technology developed by Vincent and colleagues at New England Biolabs (NEB), utilizing Escherichia Coli UvrD helicase, DNA polymerase, and single-stranded DNA-binding protein (SSB) [1]. This technique mimics the natural process of DNA replication in vivo, enabling DNA amplification in vitro under constant temperature conditions. The amplification mechanism involves the following steps. First, helicase unwinds the double-stranded DNA (dsDNA) under isothermal conditions. Then, SSB binds to the unwound single-stranded DNA (ssDNA), stabilizing it and providing a template for primer binding. Subsequently, DNA polymerase catalyzes the synthesis of double-stranded DNA. The newly synthesized dsDNA is unwound by helicase to generate single strands, which serve as templates for further cycles of DNA synthesis, resulting in exponential amplification of the target nucleic acid sequence. Compared to traditional PCR, HDA utilizes helicase and SSB to generate DNA templates under isothermal conditions, overcoming the need for thermal cycling to denature dsDNA into ssDNA. In terms of primer design and product detection, HDA is like conventional PCR. The simplicity of its principle, ease of operation, and low equipment requirements make it particularly suitable for on-site diagnostic testing, offering promising applications in this field.