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

• Published in: Biosensors & bioelectronics Vol. 86; pp. 420 - 425
• Main Authors: Tian, Bo, Qiu, Zhen, Ma, Jing, Zardán Gómez de la Torre, Teresa, Johansson, Christer, Svedlindh, Peter, Strömberg, Mattias
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 15.12.2016
• Subjects: AC susceptometer; AC susceptometry; agar gel electrophoresis; Amplification; bioassay; Brownian relaxation; Brownian relaxations; Chemical reactions; cross reaction; DNA, Viral - analysis; DNA, Viral - genetics; Equipment Design; Equipment Failure Analysis; Fluid dynamics; Fluid flow; Homogeneous detections; human; Hydrodynamic volumes; Hydrodynamics; incubation time; infection control; Isotherms; Lamps; limit of detection; Loop mediated isothermal amplifications; Loop-mediated isothermal amplification; Magnetic nano-particles; magnetic nanoparticle; Magnetic nanoparticles; Magnetometry - instrumentation; Metal Nanoparticles - chemistry; Microchemistry - instrumentation; Nanomagnetics; Nanoparticles; Nucleic Acid Amplification Techniques - instrumentation; nucleotide sequence; Oligonucleotide detection; Oligonucleotides; Polymerase chain reaction; process optimization; Proteins; Reproducibility of Results; Reverse transcription-polymerase chain reaction; RNA extraction; scanning electron microscopy; Sensitivity and Specificity; Sequence Analysis, DNA - instrumentation; serology; virus detection; virus load; Viruses; Windows (intervals); Zika fever; Zika virus; Zika Virus - genetics; Zika Virus - isolation & purification; Loop-mediated isothermal amplification; Zika virus; AC susceptometer; Brownian relaxation; Magnetic nanoparticles
• ISSN: 0956-5663; 1873-4235; 1873-4235
• DOI: 10.1016/j.bios.2016.06.085

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. •Synthetic Zika virus oligonucleotide is quantified by an AC susceptometer.•Amplicons of the LAMP reaction are attached to the magnetic nanoparticles.•Mg2P2O7 deposits onto the nanoparticles and further improves the signal.•An LOD of 1 aM oligos can be achieved with a total assay time of 27min.