Avian influenza is a serious threat to both animal and human health. To enable cutting-edge research in this field, we developed a molecular test on the basis of real-time polymerase chain reaction (real-time PCR), which detects influenza virus RNA. The test enables highly sensitive detection of influenza virus A and B strains, including H5N1, and specific identification of influenza A virus H5 subtypes. The kit was tested against a broad panel of influenza A and B subtypes and other respiratory viruses in collaboration with worldwide authoritative laboratories and shows a very high specificity and sensitivity. An internal control verifies RNA extraction as well as real-time PCR success. With this kit, rapid and reliable detection of influenza A and B viruses and identification of H5 subtypes can be achieved.