Detection of Human Hepatitis a Virus in Environmental Water by an Antigen-Capture Polymerase Chain Reaction Method

To detect hepatitis A virus (HAV) in environmental water samples, sensitive and specific methods are needed. An hemi-nested enzymatic amplification procedure was developed. When it was associated with an immuno-capture (IC) step, specificity and sensitivity were increased. The presence of a specific DNA fragment of 318 bp was detected by the analysis of the electrophoresis of the PGR products and confirmed on the autoradiogram of the Southern blot of the gel, using a labeled oligoprobe PAl selected in a very highly conserved region of the genome coding for the capsid protein VPl. The IC/PCR detected less than one infectious unit of virus in 75 µ1 sample. Eleven Seine River samples and two drinking water specimens were monitored by this assay. The IC/PCR brings a significant contribution for increasing our knowledge of the incidence of HAV on public health.