Plastic polymers for efficient DNA microarray hybridization: application to microbiological diagnostics.
Fabrication of microarray devices using traditional glass slides is not easily adaptable to integration into microfluidic systems. There is thus a need for the development of polymeric materials showing a high hybridization signal-to-background ratio, enabling sensitive detection of microbial pathogens. We have developed such plastic supports suitable for highly sensitive DNA microarray hybridizations. The proof of concept of this microarray technology was done through the detection of four human respiratory viruses that were amplified and labeled with a fluorescent dye via a sensitive reverse transcriptase PCR (RT-PCR) assay. The performance of the microarray hybridization with plastic supports made of PMMA [poly(methylmethacrylate)]-VSUVT or Zeonor 1060R was compared to that with high-quality glass slide microarrays by using both passive and microfluidic hybridization systems. Specific hybridization signal-to-background ratios comparable to that obtained with high-quality commercial glass slides were achieved with both polymeric substrates. Microarray hybridizations demonstrated an analytical sensitivity equivalent to approximately 100 viral genome copies per RT-PCR, which is at least 100-fold higher than the sensitivities of previously reported DNA hybridizations on plastic supports. Testing of these plastic polymers using a microfluidic microarray hybridization platform also showed results that were comparable to those with glass supports. In conclusion, PMMA-VSUVT and Zeonor 1060R are both suitable for highly sensitive microarray hybridizations.
Zhao
Zhengshan
Z
Centre de Recherche en Infectiologie de l'Université Laval, Centre Hospitalier Universitaire de Québec, Pavillon CHUL, 2705 Boul. Laurier, Québec, Québec, Canada.
Peytavi
Régis
R
Diaz-Quijada
Gerardo A
GA
Picard
Francois J
FJ
Huletsky
Ann
A
Leblanc
Eric
E
Frenette
Johanne
J
Boivin
Guy
G
Veres
Teodor
T
Dumoulin
Michel M
MM
Bergeron
Michel G
MG
D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010969:Plastics; D011108:Polymers; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D012680:Sensitivity and Specificity; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
D010969:Plastics; D011108:Polymers; D012367:RNA, Viral
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