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ACS Sens. 2019 Jun 28;4(6):1543-1551. doi: 10.1021/acssensors.9b00054. Epub 2019 May 16.

Nanoparticle-Based LDI-MS Immunoassay for the Multiple Diagnosis of Viral Infections.

Author information

1
Department of Bioscience and Biotechnology , National Taiwan Ocean University , Keelung 20224 , Taiwan.
2
Biomedical Engineering Research Center, Department of Electronic Engineering , Chang Gung University , Taoyuan 33302 , Taiwan.
3
Department of Nephrology , Chang Gung Memorial Hospital , Linkou, New Taipei 33305 , Taiwan.
4
Department of Materials Engineering , Ming Chi University of Technology , New Taipei City 24301 , Taiwan.
5
Department of Chemistry , Université de Montréal , Montréal , Québec H3C 3J7 , Canada.
6
Department of Biomedical Sciences, College of Medicine , Chang Gung University , Taoyuan 33302 , Taiwan.
7
Division of Pediatric Infectious Disease, Department of Pediatrics , Chang Gung Memorial Hospital , Linkou, New Taipei 33305 , Taiwan.
8
Center of Excellence for the Oceans , National Taiwan Ocean University , Keelung 20224 , Taiwan.
9
School of Pharmacy, College of Pharmacy , Kaohsiung Medical University , Kaohsiung , 80708 , Taiwan.

Abstract

Many serious public health emergencies around the globe are caused by viral epidemics. Thus, developing a reliable method for viral screening is in high demand. Multiplex assays for simultaneous detection and fast screening of high-risk pathogens are especially needed. This study employs metal nanoparticles to generate specific mass spectral signals for different RNA viruses, which enables simultaneous detection of whole viruses by laser desorption/ionization mass spectrometry (LDI-MS). We developed a nanoparticle-based sandwich immunosorbent assay as a sensing platform for the detection of viruses and viral nonstructural protein by LDI-MS. Cellulose acetate membrane (CAM) serves as the substrate for the fabrication of the sandwich immunosorbent assay with the advantages of clean mass spectra and high enrichment of analytes. Antibody-modified metal nanoparticles (Ab-MNPs; M = Au or Ag) act as metallic biocodes for the LDI-MS detection. The signal amplification readout for the virus is through the pulsed laser-induced formation of metal cluster ions ([M n]+; n = 1-3) from the Ab-MNPs which specifically bind on the CAM. Our sensing system is effective for the detection of intact viruses [Enterovirus 71 (EV71) and Japanese encephalitis virus (JEV)], nonstructural protein 1 (NS1) of Zika virus (ZIKV), EV71-spiked human serum samples, and the simultaneous detection of EV71 and ZIKV. Our probe efficiently detects EV71 in real clinical serum samples with >95% agreement with RT-qPCR results. This high-throughput LDI-MS viral detection system is simple, reliable, and high-throughput. We believe this platform has the potential to be employed for the routine screening of patients with viral infections.

KEYWORDS:

cluster ions; gold nanoparticles; laser desorption/ionization; signal amplification; simultaneous detection; viruses

PMID:
31066548
DOI:
10.1021/acssensors.9b00054

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