Silicon inhibition effects on the polymerase chain reaction: a real-time detection approach.
In the miniaturization of biochemical analysis systems, biocompatibility of the microfabricated material is a key feature to be considered. A clear insight into interactions between biological reagents and microchip materials will help to build more robust functional bio-microelectromechanical systems (BioMEMS). In the present work, a real-time polymerase chain reaction (PCR) assay was used to study the inhibition effects of silicon and native silicon oxide particles on Hepatitis B Virus (HBV) DNA PCR amplification. Silicon nanoparticles with different surface oxides were added into the PCR mixture to activate possible interactions between the silicon-related materials and the PCR reagents. Ratios of silicon nanoparticle surface area to PCR mixture volume (surface to volume ratio) varied from 4.7 to 235.5 mm2/microL. Using high speed centrifugation, the nanoparticles were pelleted to tube inner surfaces. Supernatant extracts were then used in subsequent PCR experiments. To test whether silicon materials participated in amplifications directly, in some cases, entire PCR mixture containing silicon nanoparticles were used in amplification. Fluorescence histories of PCR amplifications indicated that with the increase in surface to volume ratio, amplification efficiency decreased considerably, and within the studied ranges, the higher the particle surface oxidation, the stronger the silicon inhibition effects on PCR. Adsorption of Taq polymerase (not nucleic acid) on the silicon-related material surface was the primary cause of the inhibition phenomena and silicon did not participate in the amplification process directly.
Wang
Wei
W
Department of Engineering Mechanics, Tsinghua University, Beijing 100084, People's Republic of China. w.wang@pku.edu.cn
Wang
Hai-Bin
HB
Li
Zhi-Xin
ZX
Guo
Zeng-Yuan
ZY
D006515:Hepatitis B virus / Q000378:metabolism; D036103:Nanotechnology; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D012825:Silicon / Q000737:chemistry / Q000378:metabolism*; D013052:Spectrometry, X-Ray Emission
D012825:Silicon