The productivity of potato in Colombia is below its potential due to continued agronomical malpractices that have resulted in a high prevalence of pathogens, particularly viruses. This work presents a pilot virus seed-indexing study using the tuber sprouting test (TST) with RT-qPCR. TST was performed in certified and uncertified seed-tuber lots of potato cultivars: Diacol Capiro and Criolla Colombia. Results indicate that PYVV and PMTV are highly prevalent in all seed lots from both cultivars; PLRV, PVY, and PVB are highly prevalent in cv. Diacol Capiro, but mostly absent in cv. Criolla Colombia; PVV is almost exclusively found in cv. Criolla Colombia; while PVS and PVX are found at low prevalence in cv. Diacol Capiro. Multiple infections were common in both cultivars, and no uninfected tubers were identified. The effect of virus distribution on testing was also investigated, and a preliminary sampling method for certification of seed lots is proposed.