Comparison of hepatitis A virus concentration methods from simulated water samples using Ultrafiltration and PEG precipitation

Abstract

Objective To compare concentration methods of ultrafiltration and PEG precipitation for hepatitis A virus(HAV) in simulated water samples, and provide a reference for detection of HAV in water. Methods Quantitative HAV was inoculated in the simulated water samples. Millipore, Sartorius, 100KD and 50KD aperture microporous ultrafiltration centrifuge tubes and the final concentration of 10% polyethylene glycol (PEG) -1M sodium chloride (NaCl) precipitation method were selected to concentrate HAV. The concentration of virus was detected by fluorescence quantitative RT-PCR method. The data were statistically analyzed with SPSS21.0. Results The comparison of CT value and recovery rate shows that there were significant differences in different treatment methods (P<0.0001, P<0.0001). There were not statistically significant between ultrafiltration centrifuge tubes that they produced by different factory but with the same aperture (P=0.25, P=0.532), 50KD group had a higher recovery rate(94.57± 16.26%)than 100KD group (52.37± 14.75%) and the 10%PEG-1.0MNaCl (63.5 ± 8.45%)was in middle of them. Conclusions Both methods of ultrafiltration and PEG precipitation can be used for HAV concentration in water samples, the ultrafiltration centrifuge tube has the advantages of simple operation, less time-consuming and so on, select the appropriate aperture can improve the recovery rate of HAV virus. The HAV recovery rate of 50KD ultrafiltration centrifuge tube is higher than 100kD; PEG is more economical and practical, also it can process a large volume sample. Key words: Hepatitis A virus; viral concentration; PEG; ultrafiltration; fluorescence quantitative RT-PCR