Science Inventory

PRELIMINARY COMPARATIVE STUDY OF METHODS TO EXTRACT VIRUS FROM RAW AND PROCESSED SEWAGE SLUDGES

Citation:

RHODES, E., B. MCMINN, N. BRINKMAN, J. CASHDOLLAR, AND G. FOUT. PRELIMINARY COMPARATIVE STUDY OF METHODS TO EXTRACT VIRUS FROM RAW AND PROCESSED SEWAGE SLUDGES. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/R-07/118, 2007.

Impact/Purpose:

The objectives of this task are to:

  • Optimize a method for detecting and determining the viability of Ascaris ova in a variety of sludge types
  • Develop a Standard Operating Procedure for Ascaris ova and perform a single laboratory validation
  • Optimize a method for detecting pathogenic viruses in a variety of sludge types
  • Develop a Standard Operating Procedure for viruses and perform a single laboratory validation
  • Conduct an exposure measurement workshop on pathogens in sludg
  • e

    Description:

    Two simple virus extraction techniques were compared to an EPA standard method for detection of human enteric viruses in raw sewage sludge and class A biosolids. The techniques were used to detect both indigenous and seeded virus from a plant that distributes class A material produced by a heat drying process. Virus titers were measured using a plaque assay, a quantal assay and an integrated cell culture-reverse transcription-polymerase chain reaction (ICC-PCR) assay. The best extraction technique overall for detection of indigenous and seeded virus by plaque, quantal, and ICC-PCR assays was a simple chloroform-based technique. The detection of indigenous virus in raw sludge was similar for all three methods by the plaque assay, giving an average of 50±21 PFU/4 grams of sludge. Recovery of seeded virus from raw sludge averaged 0.7% for the EPA versus 22±17% for the other two techniques. Recovery of seeded virus from class A biosolids was similar for all techniques, averaging 95±24%. Both the quantal and the ICC-PCR assays outperformed the plaque assay for virus detection. Virus titers were generally 2->10 fold higher by these assays than by the plaque assay.



    Record Details:

    Record Type: DOCUMENT (PUBLISHED REPORT/REPORT)
    Product Published Date: 09/28/2007
    Record Last Revised: 11/03/2008
    OMB Category: Other
    Record ID: 185149

    Organization:

    U.S. ENVIRONMENTAL PROTECTION AGENCY

    OFFICE OF RESEARCH AND DEVELOPMENT

    NATIONAL EXPOSURE RESEARCH LABORATORY

    MICROBIOLOGICAL AND CHEMICAL EXPOSURE ASSESSMENT DIVISION

    BIOHAZARD ASSESSMENT RESEARCH BRANCH