Barcode technology can deliver batched information for patient healthcare. For clinical examinations, barcodes serve as reporters for labeling multiple targets, and meanwhile, facilitate improved sensitivity and specificity, thus enabling barcode as a promising alternative to traditional labels for biomarker identification and signal amplification. However, faced with the stringent claims of point-of-care (POC) bioassays, efforts are needed to advance current technologies toward rapidity, robustness, affordability, and user-friendliness. In the past decades, chemists have succeeded in delicate fabrication of the barcode libraries for encoding. Nevertheless, the decoding technologies remain poorly discussed, especially simplified decoding strategies for POC bioassays. Recent emergence of portable cartridges and miniaturized signal-recording devices has brought a promise to merge barcodes-assisted bioassay with POC testing (POCT). This review provides a comprehensive summary on barcode encoding and decoding, with emphasis on their potential use in POCT, facilitated by improved manufacturing and portable devices. Future directions of barcoded bioassays for POCT and current challenges are also presented. We anticipate that this review will be beneficial to promoting barcodes toward broad applications.