The detection of biomolecules is the key to basic molecular research, diagnostics, drug screening, and other biomedical applications. However, the existing detection techniques can only detect single classes of biomolecules, which warrant the development of a versatile biomolecule detection platform. Here, we developed a universal method for label-free detection of biomolecules via surface-enhanced Raman spectroscopy (SERS) by using sulfhydryl-modified gold nanoparticles as the substrate. The biomolecules can be adsorbed on the surface of gold nanoparticles cleaned by bromide ions to obtain initially enhanced Raman signals, and the aggregator (calcium ion) was further added to form a "hot spot", which enhanced the biomolecular signal again. Through the "two-step enhancement method", we were able to obtain fingerprints of DNA, RNA, amino acids, peptides, proteins, viruses, bacteria, and lipid molecules. This low-toxic, highly sensitive, and widely applicable technique has potential applications in biomedical research, clinical testing, and disease diagnosis and lays the foundation for the development of SERS technology in various fields.