PCR methods for the identification and detection of the...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S091200, C536S023100, C536S024300, C536S024320

Reexamination Certificate

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07097975

ABSTRACT:
Soybean rust occurs in many countries throughout Asia, Australia, Africa, and South America. The causal agents of soybean rust are two closely related fungi,Phakopsora pachyrhiziandP. meibomiae, which are differentiated based upon morphological characteristics of the telia. Determination of the nucleotide sequence of the internal transcribed spacer (ITS) region revealed greater than 99%/95% nucleotide sequence similarity among isolates of eitherP. pachyrhiziorP. meibomiae, but only 80% sequence similarity between the two species. Utilizing differences within the ITS region, four sets of PCR primers were designed specifically forP. pachyrhizi, and two sets of PCR primers were made specific toP. meibomiae. Classical and real-time fluorescent PCR assays were developed to identify and differentiate betweenP. pachyrhiziandP. meibomiae.

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Peterson et al. “Sequence comparison of rRNA ITS regions to differentiate the soybean rust speciesPhakopsora pachyrhiziandP. meibomiae”. Phytopathyology, vol. 88, No. 9, Supplement, pp. S70-71, Meeting Info. Sep. 1998.
Frederick, R., et al., “Detection and Discrimination of the Soybean Rust PathogensPhakopsora pachyrhiziandPhakopsora meibomiaeusing PCR”, (Abstract), Phytopathology, vol. 90, (6), (Supplement), p. 525, 2000.

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