A polycarbonate based surface plasmon resonance sensing cartridge for high sensitivity HBV loop-mediated isothermal amplification
Journal
Biosensors and Bioelectronics
Journal Volume
32
Journal Issue
1
Pages
89-95
Date Issued
2012
Author(s)
Abstract
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 10μl LAMP solution could be detected by SPRLAMP system in 17min even at the detection-limited concentration of 2fg/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 25min (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. ? 2011.
Subjects
Hepatitis B virus; Loop-mediated isothermal amplification; Polycarbonate; SPRLAMP sensing cartridge
SDGs
Other Subjects
Bulk solutions; Change of refractive index; Correlation coefficient; Direct sensing; DNA-template; Glass prism; Hepatitis B virus; High sensitivity; Initial concentration; Micro-reactor; On-site detection; PC-based; Sensing systems; SPRLAMP sensing cartridge; Surface plasmon resonance sensing; System response; Thermal response; Computer viruses; Plasmons; Polycarbonates; Prisms; Projectiles; Refractive index; Sensors; Viruses; Surface plasmon resonance; DNA; poly(methyl methacrylate); polycarbonate; primer DNA; article; controlled study; correlation coefficient; DNA template; hepatitis B; Hepatitis B virus; linear system; loop mediated isothermal amplification; nonhuman; refraction index; refractometer; sensitivity analysis; surface plasmon resonance; DNA, Viral; Equipment Design; Hepatitis B; Hepatitis B virus; Humans; Limit of Detection; Nucleic Acid Amplification Techniques; Polycarboxylate Cement; Refractometry; Surface Plasmon Resonance; Temperature; Hepatitis B virus
Type
journal article