Array-based nano-amplification technique was applied in detection of hepatitis E virus.
A rapid method for the detection of Hepatitis E Virus (HEV) was developed by utilizing nano-gold labeled oligonucleotide probes, silver stain enhancement and the microarray technique. The 5'-end -NH(2) modified oligonucleotide probes were immobilized on the surface of the chip base as the capture probe. The detection probe was made of the 3'-end -SH modified oligonucleotide probe and nano-gold colloid. The optimal concentrations of these two probes were determined. To test the detection sensitivity and specificity of this technique, a conservative fragment of the virus RNA was amplified by the RT-PCR/PCR one step amplification. The cDNA was hybridized with the capture probes and the detection probes on microarray. The detection signal was amplified by silver stain enhancement and could be identified by naked eyes.100 fM of amplicon could be detected out on the microarray. As the results, preparation of nano-gold was improved and faster. Development time also was shortened to 2 min. Thus, considering high efficiency, low cost, good specificity and high sensitivity, this technique is alternative for the detection of HEV.
Liu
Hui Hui
HH
State KeyLab of Virology, College of Life Sciences, Wuhan University, PR China.
Cao
Xuan
X
Yang
Yong
Y
Liu
Ming Gui
MG
Wang
Ye Fu
YF
D006046:Gold; D016752:Hepatitis E virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D049329:Nanostructures; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D044967:Serum / Q000821:virology*; D016622:Silver Staining
D015345:Oligonucleotide Probes; D006046:Gold