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Tetracycline is widely used as medicine for disease treatments and additives in animal feeding. Unfortunately, the abuse of tetracycline inevitably causes tetracycline residue in animal-origin foods. Though classical methods can detect tetracycline in high sensitivity and precision, they often rely on huge and expensive setups as well as complicated and time-consuming operations, limiting their applications in rapid and on-site detection. Here, we propose a handheld inkjet printing paper chip based smart tetracycline detector: tetracycline can be determined by inkjet printing prepared paper chip based enzyme-linked immunosorbent assay (ELISA) with the advantages of high sensitivity, excellent specificity and low cost; moreover, a smartphone based paper chip reader and application is designed for automatically determining tetracycline with simple operations, high precision and fast speed. The smart tetracycline detector with a compact size of 154 mm × 80 mm × 50 mm and self-supplied internal power can reach a rather low detection limit of ~0.05 ng/mL, as proved by practical measurements. It is believed the proposed handheld inkjet printing paper chip based smart tetracycline detector is a potential tool in antibiotic sensing for routine uses at home and on-site detection in the field.