A TaqMan real-time PCR assay for detection of Meloidogyne hapla in root galls and in soil

in Nematology
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Early detection and quantification of Meloidogyne hapla in soil is essential for effective disease management. The purpose of this study was to develop a real-time PCR assay for detection of M. hapla in soil. Primers and a TaqMan probe were designed for M. hapla detection. The assay detected M. hapla and showed no significant amplification of DNA from non-target nematodes. The assay was able to detect M. hapla in a background of plant and soil DNA. A dilution series of M. hapla eggs in soil showed a high correlation (R2=0.95, P<0.001) with Ct values. The assay could predict root-knot development in carrots by testing soils before planting. The assay could be useful for management decisions in carrot cultivation.

A TaqMan real-time PCR assay for detection of Meloidogyne hapla in root galls and in soil

in Nematology



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    Alignment including primer and probe annealing sites in the 5.8S-ITS2 region. Dots indicate conserved nucleotides and dashes indicate gaps.

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    Standard curve of Ct values plotted against the DNA concentration (ng) of Meloidogyne hapla. DNA was extracted from M. hapla eggs. Each dilution was tested in duplicate.

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    Relationship between the number of eggs in artificially infected 100 g soil and resulting Ct value. Error bars are standard deviation of three replicates.


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