PMID- 21369571
OWN - NLM
STAT- MEDLINE
DCOM- 20110721
LR  - 20110330
IS  - 1473-0189 (Electronic)
IS  - 1473-0189 (Linking)
VI  - 11
IP  - 8
DP  - 2011 Apr 21
TI  - A lab-on-a-chip device for rapid identification of avian influenza viral RNA by 
      solid-phase PCR.
PG  - 1457-63
LID - 10.1039/c0lc00528b [doi]
AB  - The endemic of Avian Influenza Virus (AIV) in Asia and epizootics in some 
      European regions have caused serious economic losses. Multiplex 
      reverse-transcriptase (RT) PCR has been developed to detect and subtype AIV. 
      However, the number of targets that can be amplified in a single run is limited 
      because of uncontrollable primer-primer interferences. In this paper, we describe 
      a lab-on-a-chip device for fast AIV screening by integrating DNA microarray-based 
      solid-phase PCR on a microfluidic chip. A simple UV cross-linking method was used 
      to immobilize the DNA probes on unmodified glass surface, which makes it 
      convenient to integrate microarray with microfluidics. This solid-phase RT-PCR 
      method combined RT amplification of extracted RNA in the liquid phase and 
      species-specific nested PCR on the solid phase. Using the developed approach, AIV 
      viruses and their subtypes were unambiguously identified by the distinct patterns 
      of amplification products. The whole process was reduced to less than 1 hour and 
      the sample volume used in the microfluidic chip was at least 10 times less than 
      in the literature. By spatially separating the primers, highly multiplexed 
      amplification can be performed in solid-phase PCR. Moreover, multiplex PCR and 
      sequence detection were done in one step, which greatly simplified the assay and 
      reduced the processing time. Furthermore, by incorporating the microarray into a 
      microchamber-based PCR chip, the sample and the reagent consumption were greatly 
      reduced, and the problems of bubble formation and solution evaporation were 
      effectively prevented. This microarray-based PCR microchip can be widely employed 
      for virus detection and effective surveillance in wild avian and in poultry 
      productions.
FAU - Sun, Yi
AU  - Sun Y
AD  - DTU Nanotech, Department of Micro- and Nanotechnology, Technical University of 
      Denmark (DTU), Kgs Lyngby, Denmark.
FAU - Dhumpa, Raghuram
AU  - Dhumpa R
FAU - Bang, Dang Duong
AU  - Bang DD
FAU - Hogberg, Jonas
AU  - Hogberg J
FAU - Handberg, Kurt
AU  - Handberg K
FAU - Wolff, Anders
AU  - Wolff A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110302
PL  - England
TA  - Lab Chip
JT  - Lab on a chip
JID - 101128948
RN  - 0 (Adhesives)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Adhesives/chemistry
MH  - Animals
MH  - Glass/chemistry
MH  - Influenza A Virus, H1N1 Subtype
MH  - Influenza A Virus, H5N1 Subtype
MH  - *Influenza A virus
MH  - *Lab-On-A-Chip Devices
MH  - Oligonucleotide Array Sequence Analysis
MH  - Polymerase Chain Reaction/*instrumentation
MH  - RNA, Viral/*analysis/*genetics
MH  - Surface Properties
MH  - Time Factors
EDAT- 2011/03/04 06:00
MHDA- 2011/07/22 06:00
CRDT- 2011/03/04 06:00
PHST- 2011/03/04 06:00 [entrez]
PHST- 2011/03/04 06:00 [pubmed]
PHST- 2011/07/22 06:00 [medline]
AID - 10.1039/c0lc00528b [doi]
PST - ppublish
SO  - Lab Chip. 2011 Apr 21;11(8):1457-63. doi: 10.1039/c0lc00528b. Epub 2011 Mar 2.