Acute　bacterial　conjunctivitis　is　a　prevalent　ocular　disease　that　can　lead　to　severe　vision　impairment　if　not　promptly　treated.　The　conventional　diagnostic　method　for　bacterial　conjunctivitis　still　relies　on　microbial　culture,　which,　although　highly　sensitive,　is　time-consuming,　labor-intensive,　and　unable　to　meet　the　demand　for　rapid　detection.　In　this　study,　we　developed　a　SERS@Au　microarray　chip　as　an　enhanced　substrate　for　collecting　the　SERS　spectra　of　conjunctivitis-associated　bacteria,　including　Escherichia　coli,　Staphylococcus　aureus,　Pseudomonas　aeruginosa,　and　Staphylococcus　epidermidis.　The　results　demonstrated　that　the　SERS@Au　microarray　chip　has　an　excellent　enhancement　effect,　reproducibility,　and　stability.　By　selecting　the　Raman　shifts　from　400　to　1　800　cm(-1)　of　pathogenic　bacteria　to　perform　an　SVM　model　and　OPLS-DA　model,　the　discrimination　accuracies　achieved　97%　and　90%,　respectively.　Detecting　spiked　tears　using　the　SERS@Au　microarrays　chip　can　quickly,　accurately,　and　conveniently　screen　pathogenic　bacteria　with　simple　culture.　Collecting　tears　for　direct　testing　is　simple,　painless,　and　non-destructive　for　patients.　Combining　the　SERS@Au　microarray　with　a　portable　Raman　spectrometer　is　suitable　for　on-site　screening　of　ophthalmic　bacterial　infections.　Moreover,　it　enables　the　detection　of　mixed　infections,　thereby　greatly　enhancing　overall　efficiency　in　diagnosis　and　providing　a　valuable　assistive　tool　for　ophthalmic　disease　screening.