Detection of potato viruses using microarray technology: towards a generic method for plant viral disease diagnosis.
Currently, most diagnostic methodology is geared towards detection of a very specific target species and often a number of assays need to be run in parallel to reach a result. The generic methods that are available for virus testing tends to give identification to the genus level only. The method described in this paper addresses this problem by exploiting a technology that has potential to test for a large number of targets in a single assay. Using the array constructed, the method was able to detect several common potato viruses (PVY, PVX, PVA, PVS) in single and mixed infections. The method was shown to be able to discriminate sequences with less than 80% sequence identity but was able to detect sequence variants with greater than 90% sequence identity. Thus the method should be useful for discriminating at the species level, but able to cope well with the intrinsic variability found within the genomes of RNA viruses. The sensitivity of the assay was found to be comparable with ELISA. The paper illustrates a significant step forward in the development of diagnostic methodologies by presenting for the first time a method that could theoretically be used not just for viruses, but for all the plant pathogens and pests that a modern diagnostic laboratory would want to test for, in a single completely generic and highly parallel format.
Boonham
N
N
Central Science Laboratory, Sand Hutton, YO41 1LZ, York, UK. n.boonham@csl.gov.uk
Walsh
K
K
Smith
P
P
Madagan
K
K
Graham
I
I
Barker
I
I
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000706:statistics & numerical data; D010935:Plant Diseases / Q000821:virology*; D017863:Potexvirus / Q000235:genetics* / Q000302:isolation & purification*; D017800:Potyvirus / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods / Q000706:statistics & numerical data
D017931:DNA Primers