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.
Highlights
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
We present a simple and homogeneous Zika virus (ZIKV) detection method that combines loop-mediated isothermal amplification (LAMP) and a magnetic AC susceptibility readout method based on the response of the magnetic nanoparticle (MNP) system, and the three step procedure is reduced to a two-step procedure
The increase in hydrodynamic volume of the MNPs resulted in a Brownian relaxation frequency shift to lower frequency which was subsequently measured by a portable AC susceptometer (Astalan et al, 2004)
Summary
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. 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. The proposed detection system can recognize 1 aM synthetic Zika virus oligonucleotide in 20% serum with a total assay time of 27 min, which can hopefully widen the detection window for Zika viremia and is promising in worldwide Zika fever control. The LAMPAC susceptometer biosensor achieves a limit of detection (LOD) of 1 aM synthetic ZIKV oligonucleotide with a total assay time of 27 min. The same sensitivity was achieved when performing the test in 20% serum samples From these results we can hopefully widen the detection window of ZIKV viremia and contribute to the Zika fever control
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