PCR inhibitor levels in concentrates of biosolid samples predicted by a new method based on excitation-emission matrix spectroscopy.
Biosolids contain a wide variety of organic contaminants that are known for their ability to inhibit PCR. During sample processing, these contaminants are coconcentrated with microorganisms. Elevated concentrations of these compounds in concentrates render samples unsuitable for molecular applications. Glycine-based elution and recovery methods have been shown to generate samples with fewer PCR inhibitory compounds than the current U.S. EPA-recommended method for pathogen recovery from biosolids. Even with glycine-based methods, PCR inhibitors still persist in concentrations that may interfere with nucleic acid amplification. This results in considerable loss of time and resources and increases the probability of false negatives. A method to estimate the degree of inhibition prior to application of molecular methods is desirable. Here we report fluorescence excitation-emission matrix (EEM) profiling as a tool for predicting levels of molecular inhibition in sample concentrates of biosolids.
Rock
Channah
C
Department of Soil, Water, and Environmental Science, University of Arizona, Maricopa Agricultural Center, Maricopa, Arizona 85138, USA.
Alum
Absar
A
Abbaszadegan
Morteza
M
D004783:Environmental Microbiology; D004791:Enzyme Inhibitors / Q000032:analysis*; D008828:Microbiological Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013057:Spectrum Analysis / Q000379:methods*
D004791:Enzyme Inhibitors