Pre-analytical and analytical procedures for the detection of enteric viruses and enterovirus in water samples

A one-step centrifugal ultrafiltration method to concentrate enteric viruses from wastewater

“…Currently, the most common concentration method is virus adsorption-elution (VIRADEL) using electropositive or electronegative microporous membrane filters, such as Virosorb 1MDS and NanoCeram filters (Ikner et al, 2012;Pang et al, 2012;Cashdollar and Wymer, 2013). The process involves the absorption of viral particles to the membrane through ionic interaction followed by elution with pH adjustment.…”

“…Pang et al reported a virus concentration method that used NanoCeram disc filtration, beef extract elution and flocculation based on VIRADEL (Pang et al, 2012). This method has been used to concentrate viruses in both wastewater (Qiu et al, 2015) and surface water (Pang et al, 2012).…”

“…This method has been used to concentrate viruses in both wastewater (Qiu et al, 2015) and surface water (Pang et al, 2012). However, there are limitations with this method, such as a large sample volume to be processed, the loss of virions due to multiple processing steps, long processing time and limited sample throughput.…”

“…In this study, a simple and rapid one-step centrifugal ultrafiltration method was developed using the Centricon Plus-70 centrifugal ultrafilter (30-kDa MWCO, Millipore) to concentrate human enteric viruses from wastewater. In order to determine the effectiveness of the one-step ultrafiltration method, VIRADEL developed by Pang et al (2012) was used in parallel to concentrate virus from the same water samples. Real-time quantitative PCR (rt-qPCR) and cell culture were used to quantify the amount of virus/free viral nucleic acids and infectious virions respectively after each concentration method.…”

“…A total of 12 samples were collected by directly grabbing from the wastewater flow before UV treatment (pre-UV) and post-UV treatment respectively during the study. Ten liters of grab water samples were processed using VIRADEL as previously described (Pang et al, 2012;Qiu et al, 2015). The centrifugal ultrafiltration was used to concentrate virus from 100 mL wastewater sample using the Centricon Plus-70 filter according to the Manufacturer's instructions, except for the pre-rinse step.…”