A procedure using Rotylenchulus reniformis nucleotide sequences to quantitatively measure plant-parasitic nematode infestation levels using metagenomic DNA isolated directly from soil

Molecular diagnostic tests have been developed and utilized to diagnose and to confirm diagnoses of many plant-parasitic nematodes. We evaluate the potential of a qPCR assay to detect and quantify Rotylenchulus reniformisin Mississippi directly from soil. A novel pipeline utilizing multiple databases containing nematode DNA and EST sequences was developed to aid in the selection of R. reniformis primers used in a PCR and qPCR assays. In vitro testing showed that the primers and probes developed from the novel pipeline for the qPCR assays could accurately detect the presence of R. reniformis. Subsequent testing resulted in a trend of increasing observed number of R. reniformis resulting in increasing estimates by qPCR.