Home › Microbiology › EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. II. Total Culturable Virus Assay
Microbiology JoVE (Open Access) Citable · DOI

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. II. Total Culturable Virus Assay

DOI: 10.3791/52437-v
What you'll learn
  • ✓ Perform filter elution and organic flocculation to concentrate viruses from water samples
  • ✓ Execute a quantal assay using Buffalo Green Monkey kidney cells to measure culturable viruses
  • ✓ Apply EPA Method 1615 standardized protocols for infectious virus detection in environmental waters
Protocol

Here we present a procedure to measure total culturable viruses using the Buffalo Green Monkey kidney cell line. The procedure provides a standardized tool for measuring the occurrence of infectious viruses in environmental and drinking waters.

Difficulty
advanced
Total time
~5–7 days (includes virus propagation, sample concentration, cell culture assay, and cytopathic effect observation)
Model organism
Buffalo Green Monkey kidney cells
Biosafety
BSL-2

Steps

1
Perform filter elution procedure on water samples

Elute concentrated viruses from filter membranes using standardized elution buffer to recover viral particles from environmental water samples.

▶ 02:32
2
Concentrate viruses using organic flocculation

Apply organic flocculation concentration method to further concentrate eluted viral particles prior to cell culture assay.

▶ 03:33
3
Conduct total culturable virus quantal assay

Inoculate serial dilutions of concentrated virus onto Buffalo Green Monkey kidney cells and observe cytopathic effects to quantify infectious viral titer.

▶ 06:03
4
Evaluate poliovirus recovery from water samples

Assess mean poliovirus recovery percentage from ground and reagent-grade water to validate assay performance.

▶ 09:26
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