Hepatitis A virus detection in wastewater by PCR and hybridization

Hepatitis A virus is a member of the Picornaviridae family and is a principal agent of acute hepatitis worldwide, causing from mild to severe illness. Although the incidence of hepatitis A is in decline, the risk of this disease is still high in the Mediterranean area. Detection of hepatitis A in the environment is difficult because this virus needs a prolonged incubation in cell culture, therefore we used an antigen capture PCR (AC-PCR) followed by a hybridization on membrane to identify HAV in wastewater samples. The raw sewage, concentrated by ultrafiltration, showed 8 positive samples out of 10 (80%), while after the oxidation step of the sewage, 2 out of 10 (20%) and 3 out of 10 (30%) were found positive respectively after concentration by electronegative (HAWP Millipore) and electropositive (1MDS Cuno-Div.) membranes. In the final effluent the positivity was 1 out of 10 (10%) for the electronegative membranes and 3 out of 10 (30%) for the electropositive membranes. Our results indicate: i) the possibility of HAV to cross the wastewater treatment plant and contaminate water and food (such as mussels); ii) PCR-hybridization as a rapid method for HAV identification in the environment.