Recent outbreaks of avian influenza (AI) occurring in Europe, in the Americas, in Asia and in Africa have provided field evidence of how challenging the control of this infection can be, particularly in densely populated poultry areas or in areas where free-range rural village poultry and backyard flocks are present. In these areas, laboratory testing is mainly applied to trace viral circulation in a given area or in a susceptible population to implement an early warning system, rather than to diagnose the presence of the virus in a diseased flock or animal. This implies the use of rapid, sensitive and possibly cost-effective laboratory tests adaptable to very high throughputs. As a consequence, new diagnostic approaches and technologies have been increasingly developed and applied. Molecular biology and biotechnology are providing important and precious contributions to the field of AI diagnosis, making extremely rapid, specific and sensitive techniques available. However, the use of some of these technologies is still limited, due to their costs and to the requirement of advanced technical and scientific expertise. Therefore, more conventional and well-established techniques, should not be abandoned but rather reconsidered and improved or modified.