Flexible microarray construction and fast DNA hybridization conducted on a microfluidic chip for greenhouse plant fungal pathogen detection

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Author(s)

Detail(s)

Original languageEnglish
Pages (from-to)10509-10516
Journal / PublicationJournal of Agricultural and Food Chemistry
Volume55
Issue number26
Publication statusPublished - 26 Dec 2007
Externally publishedYes

Abstract

This study employed a microfluidic method in which probe creation does not require pin-spotting and fast hybridization is conducted on the same microarray chip for the detection of three greenhouse pathogens (Botrytis cinerea, Didymella bryoniae, and Botrytis squamosa). In this method, 16 oligonucleotide probe line arrays were created on a glass substrate by a microfluidic printing method. Then, low amounts of the DNA samples (1 fmol of oligonucelotides or 1.4 ng of PCR products) were introduced into the microchannels that were orthogonal to these probe lines. The hybridizations of 16 samples (21-mer complementary oligonuleotides and ∼260 bp PCR products) were fulfilled at the channel-probe line intersections and in a short time (minutes). The optimization of probe immobilization and sample hybridization are described in detail. The method successfully detected and discriminated between two 260 bp PCR products with a one-base-pair difference from closely related greenhouse plant fungal pathogens (B. cinerea and B. squamosa). © 2007 American Chemical Society.

Research Area(s)

  • DNA hybridization, Fungal pathogen, Microfluidic microarray, Single base-pair difference discrimination

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