To further reduce the size of a microfluidic detection chip and the sample consumption and to shorten the chip manufacturing cycle, an all-fiber SPR detection multichannel microfluidic chip was proposed and demonstrated in this paper. The microfluidic channel of the proposed chip was provided by the air channel of a double side-hole fiber, the detection unit was fabricated using a dumbbell fiber with a fiber core exposed to air, and the sensing probe was composed and packaged by fiber micro-processing technology. The internal double channels of the fiber constructed from double side-hole and dumbbell fibers can realize dual channel detection based on space division multiplexing. 30 nm silver and 50 nm gold films were respectively coated on the left and right sides of the dumbbell fiber, which can realize the dual channel simultaneous detection based on wavelength division multiplexing. We employed the proposed microfluidic chip to detect immunoglobulin G and dopamine molecules, where the average sensitivity is 0.252 nm (mg mL⁻¹)⁻¹ and 0.061 nm (μg mL⁻¹)⁻¹, and the LOD is 0.397 mg mL⁻¹ and 1.639 μg mL⁻¹, respectively. The microfluidic channel and detection unit of all-fiber multi-channel SPR detection microfluidic chip are provided by a soft and flexible fiber, which is compact in structure, flexible in fabrication and short in manufacturing cycle, making it possible for the microfluidic chip to enter the human body for detection and enabling a new approach for the fabrication of wearable detection microfluidic devices. This provides a new idea for the development of microfluidic chips.