Puumala and Dobrava Hantaviruses

Throughout the world nearly 150.000 persons are infected with hantaviruses each year. Hantaviruses are divided into two groups based on their human pathogenicity: Old World hantaviruses causing hemorrhagic fever with renal syndrome (HFRS) and New World hantaviruses causing hantavirus pulmonary syndrome (HPS). HFRS is an acute disease characterised by vascular bleeding and kidney damage and in most cases it is not lethal. Mild forms of HFRS are usually caused by Puumala virus (PUUV) while the most severe forms of HFRS in Europe are caused by Dobrava-Belgrade virus (DOBV). Therefore sensitive and specific molecular detection is essential for the timely diagnosis of hantavirus infections. In a typical laboratory context we present the creation of nested degenerate primers for detection of S and M segment of DOBV and M segment of PUUV optimisation of various PCR components and cycling parameters using both external and internal primers. As a result this book provides a resource for anyone interested in conducting similar experiments and procedures.