We present a theoretical framework for the analysis of the statistical properties of thermal fluctuations on a lossy transmission line. A quantization scheme of the electrical signals in the transmission line is formulated. We discuss two applications in detail. Noise spectra at finite temperature for voltage and current are shown to deviate significantly from the Johnson-Nyquist limit, and they depend on the position on the transmission line. We analyze the spontaneous emission, at low temperature, of a Rydberg atom and its resonant enhancement due to vacuum fluctuations in a capacitively coupled transmission line. The theory can also be applied to study the performance of microscale and nanoscale devices, including high-resolution sensors and quantum information processors.