Robust PCR assay for early detection of Microcystin-Producing Cyanobacteria in Surface Water Resources

Background

The contamination of surface water resources by toxin producing cyanobacterial (blue green algae) blooms is a global problem. 

A rapid and reliable early-warning monitoring system is required for local drinking water sources. This allows actions to mitigate bloom formation and/or prevent contaminated water from entering drinking water treatment plants.

Description

A practical, robust and sensitive PCR test for detecting potentially microcystin-producing cyanobacteria in surface water samples is provided:

  • Allows identification of the cyanobacterial genus (Microcystis, Planktothrix and Dolichospermum).
  • Utilizes a single ~100 bp amplicon within the mcyB gene combined with target-specific probes or melting curve analysis.
  • Allows copy number quantification of the mcyB gene.
  • Utilizes easy-to-use disposable dry chemistry chips (Fig 1).
  • The quantification range is 5 × 100 to 1 × 107 copies of mcyB gene per reaction.

Application Areas and Potential End Users

Monitoring of drinking or recreational water samples prior to, as well as during full-scale blooms. Potential end users include urban water plants, drinking water and environmental authorities and water testing laboratories. 

The IP is available for licensing by test developers and testing service providers from the TTO of the University of Turku (UTU)

Title:  METHOD AND PRIMERS FOR THE DETECTION OF MICROCYSTIN-PRODUCING TOXIC CYANOBACTERIA  

Patent: EP2776584
Technology Readiness Level (TRL): 6

Status: Practical dry chemistry all-in-one assays have been produced and the testing concept has been validated e.g. at local drinking water treatment plants (Figure 2). 

The current version of the test utilizes a disposable assay chip (Figure 1) and an integrated time-resolved fluorescence PCR device. The short (~100 bp) amplicon and simplified assay designs allows the assay to be adapted to any nucleic acid assay platform selected by the licensee. 

Figure 1. Disposable dry chemistry assay chips
Figure 1. Disposable dry chemistry assay chips

 

Figure 2. Positive correlation between total microcystin concentrations (HPLC-FLD) and mcyB copy numbers in lake and river water samples. (Savela et al. 2014.)
Figure 2. Positive correlation between total microcystin concentrations (HPLC-FLD) and mcyB copy numbers in lake and river water samples. (Savela et al. 2014.)