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Diagnostic Values and Utility of Immunological, Morphological, and Molecular Methods for In Planta Detection of Phytophthora ramorum

September 2007 , Volume 97 , Number  9
Pages  1,119 - 1,129

L. F. F. Kox, I. R. van Brouwershaven, B. T. L. H. van de Vossenberg, H. E. van den Beld, P. J. M. Bonants, and J. de Gruyter

First, second, third, fourth, and sixth authors: Plantenziektenkundige Dienst, P.O. Box 9102, 6700 HC Wageningen, The Netherlands; and fifth author: Plant Research International B.V., P.O. Box 16, 6700 AA Wageningen, The Netherlands.

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Accepted for publication 18 April 2007.

In this study, six methods for the detection of Phytophthora ramorum in planta were compared using naturally infested rhododendron plant material. The methods included two immunological methods, one an enzyme-linked immunosorbent assay (ELISA) and the other using a lateral flow format (LFD). Three molecular tests based on the polymerase chain reaction (PCR) using TaqMan chemistry also were assessed, including two assays designed for specific detection of P. ramorum and one designed for genus-level detection of Phytophthora. Isolation followed by morphological identification also was assessed. The diagnostic values of each of the methods, evaluated based on diagnostic sensitivity, diagnostic specificity, positive predictive value, and negative predictive value, were calculated based upon the test results from 148 field samples. The “gold standard” used for the calculations was the final diagnosis, which was based on either a positive PCR result or successful isolation of P. ramorum. The Phytophthora spp. TaqMan PCR, ELISA, and LFD had higher sensitivities than the P. ramorum-specific methods, which make them useful as prescreening methods, where positive results must be confirmed by PCR or isolation. The article discusses practical advantages and disadvantages of each of the methods and how they are valuable in the diagnostic process, according to the circumstances of use (that is, diagnosis or surveillance) and in relation to the prevalence of P. ramorum infestation in the population to be tested.

Additional keywords: analytical sensitivity, analytical specificity, DNA isolation, sudden oak death.

The American Phytopathological Society, 2007