Polyethylene glycol precipitation for recovery of pathogenic viruses, including hepatitis A virus and human rotavirus, from oyster, water, and sediment samples

Abstract

Polyethylene glycol 6000 precipitation was found to be an effective concentration method that enhanced the chances for detecting human virus pathogens in environmental samples. Percent recoveries from eluates of fresh and estuarine waters with 8% polyethylene glycol 6000 averaged 86 for hepatitis A virus, 77 for human rotavirus Wa, 87 for simian rotavirus SAll, and 68 for poliovirus. Percent recoveries of 97, 40, 97 and 105, respectively, for the same viruses were obtained from oyster eluates by the same procedure. Percent recoveries of 97 for hepatitis A virus and 78 for human rotavirus Wa were obtained from sediment eluates containing 2 M NaNO3 with a final concentration of 15% polyethylene glycol 6000. The polyethylene glycol method was shown to be more effective than the organic flocculation method for recovery of hepatitis A virus and rotaviruses Wa and SAll, but not of poliovirus 1 in laboratory studies. In field trials, hepatitis A virus or rotavirus or both were recovered from 12 of 18 eluates by polyethylene glycol, compared with recovery from 9 of 18 eluates by organic flocculation from fresh and estuarine waters subject to pollution.

Lethal murine infection model for human respiratory disease-associated Pteropine orthoreovirus

“…The quail fibrosarcoma QT6 cell line was grown in DMEM supplemented with 10% fetal calf serum. PRV strain MB (Yamanaka et al, 2014) was amplified in L929 cells and approximately 10-fold concentrated virus stock was generated using a 10% polyethylene glycol concentration method as described elsewhere (Lewis and Metcalf, 1988). Infectious virus titer was determined by a plaque assay (Kawagishi et al, 2016).…”

Cellulose-based virus-retentive filters: a review

“…Hence, Tables 4 and 5 display a wide range of recovery yields. In practical tests aimed at the detection of naturallyoccurring viruses in field, large-volume water samples, the filtration (adsorption-elution) step is usually a primary concentration step which is followed by a secondary one to reach the virus detection threshold, e.g., organic flocculation or PEG precipitation (Lewis and Metcalf 1988). The recovery yield of such two-step concentration processes was evaluated in several studies using seeded or naturally contaminated water samples (Tables 4, 5).…”

New tools for the study and direct surveillance of viral pathogens in water

“…Positively charged filters [23] and glass wool [24] based methods are still among the best possibilities. Sampling large volumes requires a two-step concentration procedure, with polyethylene glycol precipitation [25] and ultrafiltration [26] Virus detection in water Bosch et al 297…”

“…as preferred procedures for reconcentration of the primary eluates. Additionally, PEG [25] as well as lyophilisation [3] may be used for direct virus concentration in heavily polluted medium size samples, for example, sewage, having this latter method the added advantage of removing substances inhibitory to RT-PCR enzymes if this method is employed for virus detection [27].…”