A polycarbonate based surface plasmon resonance sensing cartridge for high sensitivity HBV loop-mediated isothermal amplification.
In this study, we report a simple, low-cost surface plasmon resonance (SPR)-sensing cartridge based on a loop-mediated isothermal amplification (LAMP) method for the on-site detection of the hepatitis B virus (HBV). For LAMP detection, a SPR based LAMP sensing system (SPRLAMP) was constructed, including a novel SPRLAMP sensing cartridge integrating a polymethyl methacrylate (PMMA) micro-reactor with a polycarbonate (PC)-based prism coated with a 50 nm Au film. First, we found that the change of refractive index of the bulk solution was approximately 0.0011 refractive index (RI) units after LAMP reaction. The PC-based prism's linearity and thermal responses were compared to those of a traditional glass prism to show that a PC-based prism can be used for SPR measurement. Finally, the HBV template mixed in the 10 μl LAMP solution could be detected by SPRLAMP system in 17 min even at the detection-limited concentration of 2 fg/ml. We also analyzed the correlation coefficients between the initial concentrations of HBV DNA templates and the system response (ΔRU) at varying amplification times to establish an optimal amplification time endpoint of 25 min (R(2)=0.98). In conclusion, the LAMP reaction could be detected with the SPRLAMP sensing cartridge based on direct sensing of the bulk refractive index.
Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan, ROC.
Tsung-Liang
TL
Chuang
Shih-Chung
SC
Wei
Szu-Yuan
SY
Lee
Chii-Wann
CW
Lin
D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D004867:Equipment Design; D006509:Hepatitis B / Q000175:diagnosis; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D011076:Polycarboxylate Cement / Q000737:chemistry*; D012031:Refractometry; D020349:Surface Plasmon Resonance / Q000191:economics / Q000295:instrumentation*; D013696:Temperature
D004279:DNA, Viral; D011076:Polycarboxylate Cement; C005506:polycarbonate