A 3D porous graphene structure was directly induced by CO2 laser from the surface of Kapton tape (carbon source) supported by polyethylene terephthalate (PET) laminating film. A highly flexible laser-induced porous graphene (LI-PGr) electrode was then fabricated via a facile one-step method without reagent and solvent in a procedure that required no stencil mask. The method makes pattern design easy, and production cost-effective and scalable. We investigated the performance of the LI-PGr electrode for the detection of methamphetamine (MA) on household surfaces and in biological fluids. The material properties and morphology of LI-PGr were analysed by scanning electron microscopy (SEM), energy dispersive x-ray (EDX) and Raman spectroscopy. The LI-PGr electrode was used as the detector in a portable electrochemical sensor, which exhibited a linear range from 1.00 to 30.0 µg mL−1 and a detection limit of 0.31 µg mL−1. Reproducibility was good (relative standard deviation of 2.50% at 10.0 µg mL−1; n = 10) and anti-interference was excellent. The sensor showed good precision and successfully determined MA on household surfaces and in saliva samples.