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Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses
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Published on: December 3, 2011

Performance of forty-one microbial source tracking methods: a twenty-seven lab evaluation study.

Alexandria B Boehm1, Laurie C Van De Werfhorst, John F Griffith

  • 1Environmental and Water Studies, Department of Civil and Environmental Engineering, Stanford University, Stanford, CA 94305, USA.

Water Research
|July 25, 2013
PubMed
Summary

This study evaluated 41 microbial source tracking (MST) methods across 27 labs. The best performing assays for human, ruminant, gull, and pig sources showed high sensitivity and specificity, though further testing is needed for portability.

Keywords:
BacteroidalesEnterococciFecal pollutionMicrobial source tracking

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Area of Science:

  • Environmental Microbiology
  • Molecular Biology
  • Water Quality Assessment

Background:

  • Numerous microbial source tracking (MST) methods have been developed.
  • Limited inter-laboratory validation and testing with diverse fecal samples exist for many MST methods.

Purpose of the Study:

  • To evaluate the specificity and sensitivity of 41 MST methodologies.
  • To assess the performance of MST assays targeting various animal and human fecal sources across multiple laboratories.

Main Methods:

  • Analysis of data from 27 laboratories testing 64 blind fecal samples (singletons, doubletons, diluted samples).
  • Evaluation of 41 MST methodologies targeting human, cow, ruminant, dog, gull, pig, horse, and sheep.
  • Assessment of assay performance using both binary (presence/absence) and quantitative (abundance) analyses.

Main Results:

  • Several assays demonstrated high sensitivity and specificity (>80%) in binary analysis, including HF183 (human), Rum2Bac (ruminant), CowM2/M3 (cow), BacCan (dog), LeeSeaGull (gull), and pig-specific assays.
  • Quantitative analysis revealed HF183Taqman and BacH (human), Rum2Bac and BacR (ruminant), LeeSeaGull (gull), and Pig2Bac (pig) as top performers.
  • No cow or dog-specific assays met quantitative criteria; some assays with good binary performance lacked sufficient sensitivity in quantitative analysis.

Conclusions:

  • The study identified robust MST assays for specific fecal sources, particularly for human, ruminant, gull, and pig.
  • Assay portability requires further investigation as some top performers were only tested in one laboratory.
  • Development of frameworks for fecal source allocation and risk assessment, alongside field testing, is crucial for practical application.