A simple method for concentrating and detecting viruses in wastewater

A simple method has been developed for routine analysis of sewage and sewage effluents for detecting viruses using adsorption at pH 3 on a 0·45 μm 47 mm diameter membrane filter and elution at pH 8. It was tested on viruses added to autoclaved sewage. Homogenizing the sample for 4 min in a Waring blender and clarification by centrifugation at 1800 g and later at 9230 g facilitated easy filtration without any loss of virus. Retention of the eluant for 30 min on the millipore membrane and then elution in situ under suction provided a sterile eluate with 100 per cent recovery of viruses.

Viruses added to fecal suspensions with 600 mg I⁻¹ BOD were completely recovered when the sample pH was adjusted to 3 and its salt concentration increased by adding 1200 mg l⁻¹ of Mg²⁺ as the chloride. This procedure eliminated the need for passing the samples through ion exchange resins for removing membrane coating components. In a 1 yr programme of monitoring of raw sewage from a middle income group community in Nagpur, a maximum of 3150 PFU/1 during monsoon and 11575 PFU/1 during winter was obtained.

High efficiency and reproducibility of the method allowed the use of sample volumes of 40 ml of raw sewage and 320 ml of treated effluent for the detection of viruses.