A 3.5-year-old Yorkshire Terrier was evaluated for anorexia and vomiting; infection with Mycobacterium tuberculosis was diagnosed by use of histology, bacteriologic culture, and polymerase chain reaction (PCR) assay on various tissues. The dog was living with a human with an established M. tuberculosis infection. Findings were unique in that diagnosis of M. tuberculosis infection was obtained via PCR techniques, and isolates from the owner and dog were matched via restriction fragment length polymorphism fingerprinting. Dogs infected with M. tuberculosis from humans are most commonly infected via the respiratory tract. Clinical signs in dogs are variable and depend on the integrity of the immune system and the degree of dissemination. Diagnosis can often be obtained through histopathology and bacteriologic culture; additional diagnostic techniques are also available. Treatment of a dog with confirmed M. tuberculosis infection is controversial, and at least 6 months of multidrug treatment is required.