We present the first study of the variable star populations in the isolated dwarf spheroidal galaxies (dSphs) Cetus and Tucana. Based on Hubble Space Telescope images obtained with the Advanced Camera for Surveys in the F475W and F814W bands, we identified 180 and 371 variables in Cetus and Tucana, respectively. The vast majority are RR Lyrae stars. In Cetus, we also found three anomalous Cepheids (ACs), four candidate binaries and one candidate long-period variable (LPV), while six ACs and seven LPV candidates were found in Tucana. Of the RR Lyrae stars, 147 were identified as fundamental mode (RRab) and only eight as first-overtone mode (RRc) in Cetus, with mean periods of 0.614 and 0.363 day, respectively. In Tucana, we found 216 RRab and 82 RRc giving mean periods of 0.604 and 0.353 day. These values place both galaxies in the so-called Oosterhoff Gap, as is generally the case for dSph. We calculated the distance modulus to both galaxies using different approaches based on the properties of RRab and RRc, namely, the luminosity-metallicity and period-luminosity-metallicity relations, and found values in excellent agreement with previous estimates using independent methods: (m - M)0,Cet = 24.46 ± 0.12 and (m - M)0,Tuc = 24.74 ± 0.12, corresponding to 780 ± 40 kpc and 890 ± 50 kpc. We also found numerous RR Lyrae variables pulsating in both modes simultaneously (RRd): 17 in Cetus and 60 in Tucana. Tucana is, after Fornax, the second dSph in which such a large fraction of RRd (~17%) has been observed. We provide the photometry and pulsation parameters for all the variables, and compare the latter with values from the literature for well studied dSph of the Local Group and Galactic globular clusters. The parallel WFPC2 fields were also searched for variables, as they lie well within the tidal radius of Cetus, and at its limit in the case of Tucana. No variables were found in the latter, while 15 were discovered in the outer field of Cetus (11 RRab, three RRc, and one RRd), even though the lower signal-to-noise ratio of the observations did not allow us to measure their periods accurately. We provide their coordinates and approximate properties for completeness. Based on observations made with the NASA/ESA Hubble Space Telescope, obtained at the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS5-26555. These observations are associated with program 10505.