Leptospirosis is caused by pathogenic bacteria called leptospires that are transmitted directly or indirectly from animals to humans. It occurs worldwide but is most common in tropical and subtropical areas. It is a potentially serious but treatable disease. Its symptoms may mimic those of a number of other unrelated infections such as influenza, meningitis, hepatitis, dengue, or other viral hemorrhagic fevers. The spectrum of the disease is extremely wide, ranging from subclinical infection to a severe syndrome of multiorgan infection with high mortality. Laboratory diagnosis tests are not always available, especially in developing countries. Numerous tests have been developed, but availability of appropriate laboratory support is still a problem. Direct observation of leptospires by darkfield microscopy is unreliable and not recommended. Isolation of leptospires can take up to months and does not contribute to early diagnosis. Diagnosis is usually performed by serology; enzyme-linked immunosorbent assay and the microscopic agglutination tests are the laboratory methods generally used, rapid tests are also available. Limitation of serology is that antibodies are lacking at the acute phase of the disease. In recent years, several real-time polymerase chain reaction assays have been described. These can confirm the diagnosis in the early phase of the disease prior to antibody titers are at detectable levels, but molecular testing is not available in restricted resources areas.