The stretchable sensor has been considered as the most important component in a wearable device. However, it is still a great challenge to develop a highly sensitive textile-based strain sensor with good flexibility, excellent skin affinity, and large dynamic range. Herein, polypyrrole (PPy) was immobilized on a stretchable textile knitted by polyester and spandex via low-temperature interfacial polymerization to prepare a conductive strain sensor for human motion and respiration measurements. Scanning electron microscopy, Fourier transform infrared spectrometry, and thermal gravimetric data verify that a thin layer of PPy has been successfully coated on the textile with a high density and very uniform distribution. The resistance of the as-prepared textile is 21.25 Ω/cm2 and the PPy-coated textile could be used as an electric conductor to light up a LED lamp. Moreover, the textile could tolerate folding at an angle of 180° and 500 times of bending-twisting cycles without significant changes on its resistance. A negative correlation between the resistance change and the applied strain is observed for the textile-based sensor in the strain ranging from 0 to 71% with the gauge factor of −0.46. After more than 200 cycles of stretching-releasing under the strain of 26%, there is no obvious alteration on the sensing responses. The sensors were attached on volunteers’ body or clothes for the real-time measurement of human motions and respiration, demonstrating that the textile-based sensor could sensitively detect finger, elbow, and knee bending and differentiate deep, normal, and fast breath. This work may provide an approach to uniform and dense coating conductive polymers on textiles for highly sensitive and stretchable sensors, which possess great potentials in practical applications for real-time monitoring human motions and physiological signs.