Attomolar Zika virus oligonucleotide detection based on loop-mediated isothermal amplification and AC susceptometry

Biosens Bioelectron. 2016 Dec 15:86:420-425. doi: 10.1016/j.bios.2016.06.085. Epub 2016 Jun 29.

Abstract

Because of the serological cross-reactivity among the flaviviruses, molecular detection methods, such as reverse-transcription polymerase chain reaction (RT-PCR), play an important role in the recent Zika outbreak. However, due to the limited sensitivity, the detection window of RT-PCR for Zika viremia is only about one week after symptom onset. By combining loop-mediated isothermal amplification (LAMP) and AC susceptometry, we demonstrate a rapid and homogeneous detection system for the Zika virus oligonucleotide. Streptavidin-magnetic nanoparticles (streptavidin-MNPs) are premixed with LAMP reagents including the analyte and biotinylated primers, and their hydrodynamic volumes are dramatically increased after a successful LAMP reaction. Analyzed by a portable AC susceptometer, the changes of the hydrodynamic volume are probed as Brownian relaxation frequency shifts, which can be used to quantify the Zika virus oligonucleotide. The proposed detection system can recognize 1 aM synthetic Zika virus oligonucleotide in 20% serum with a total assay time of 27min, which can hopefully widen the detection window for Zika viremia and is therefore promising in worldwide Zika fever control.

Keywords: AC susceptometer; Brownian relaxation; Loop-mediated isothermal amplification; Magnetic nanoparticles; Zika virus.

MeSH terms

  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Equipment Design
  • Equipment Failure Analysis
  • Magnetometry / instrumentation*
  • Metal Nanoparticles / chemistry
  • Microchemistry / instrumentation
  • Nucleic Acid Amplification Techniques / instrumentation*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / instrumentation*
  • Zika Virus / genetics*
  • Zika Virus / isolation & purification*

Substances

  • DNA, Viral