Rapid and Ultrasensitive Quantification of Multiplex Respiratory Tract Infection Pathogen via Lateral Flow Microarray based on SERS Nanotags

Abstract

Respiratory tract infections (RTIs) are severe acute infectious diseases, which require the timely and accurate identification of the pathogens involved so that the individual treatment plan can be selected, including optimized use of antibiotics. However, high throughput and ultrasensitive quantification of multiple nucleic acids is a challenge in a point of care testing (POCT) device. Methods: Herein, we developed a 2×3 microarray on a lateral flow strip with surface enhanced Raman scattering (SERS) nanotags encoding the nucleic acids of 11 common RTI pathogens. On account of the signal magnification of encoded SERS nanotags in addition to the high surface area to volume ratio of the nitrocellulose (NC) membrane, rapid quantification of the 11 pathogens with a broad linear dynamic range (LDR) and ultra-high sensitivity was achieved on one lateral flow microarray. Results: The limit of detection (LOD) for influenza A, parainfluenza 1, parainfluenza 3, respiratory syncytial virus, coxiella burnetii, legionella pneumophila, influenza B, parainfluenza 2, adenovirus, chlamydophila pneumoniae, and mycoplasma pneumoniae were calculated to be 0.031 pM, 0.030 pM, 0.038 pM, 0.038 pM, 0.040 pM, 0.039 pM, 0.035 pM, 0.032 pM, 0.040 pM, 0.039 pM, and 0.041 pM, respectively. The LDR of measurement of the target nucleic acids of the eleven RTI pathogens were 1 pM-50 nM, which span 5 orders of magnitude. Conclusions: We anticipate this novel approach could be widely adopted in the early and precise diagnosis of RTI and other diseases.

PRADA: Portable Reusable Accurate Diagnostics with nanostar Antennas for multiplexed biomarker screening

“…The corresponding sensitivity curve obtained from the SERS measurements was fitted using the four‐parameter logistic (4PL) function (Table S1) showing that the Raman intensity increased in a logarithmic manner with increasing concentrations of cTnI in the range of 0.02 to 5,000 ng/ml (Figure 3b). This 4PL function has been shown previously to have a robust fit to plasmonic and SERS‐based biosensors 5,44 . However, the SERS intensity at 1325 cm −1 was linear at low concentrations of cTnI in the quantification region (Figure 3c).…”

“…This 4PL function has been shown previously to have a robust fit to plasmonic and SERS-based biosensors. 5,44 However, the SERS intensity at 1325 cm −1 was linear at low concentrations of cTnI in the quantification region ( Figure 3c). The LOD of cTnI was estimated to be 0.0055 ng/ml from this fit; all parameters for LOD calculation are provided in Table S2.…”

A materials-science perspective on tackling COVID-19

“…For example, an Fe–Au nanoparticle-coupling approach, combining PCR amplification with the quantitative measurement of Au nanoparticles using inductively coupled plasma mass spectrometry 129 , which was originally developed for the detection of circulating tumour DNA, could also be applied for the sensitive and specific detection of SARS-CoV-2 RNA. Microarrays, which have been developed for the precise control and manipulation of fluids, combine sample preparation, reaction, separation and detection on one chip of a few square centimetres 130 , and could be used in combination with surface-enhanced Raman spectroscopy for rapid virus detection using nanoparticle aggregates or arrays that can generate surface-enhanced Raman spectroscopy signals 131 , 132 . Low-cost point-of-care devices, designed with microfluidics, could also be adapted for viral detection, requiring only small sample volumes.…”

Surface-Enhanced Raman Scattering-Based Lateral-Flow Immunoassay

“…Recently, core–shell-type metal nanoparticles have also been widely used for designing SERS probes. By combining the high stability and monodispersity of gold and the large enhancement factor of silver, Au@Ag [ 27 , 45 , 46 , 47 , 48 , 49 , 50 , 51 ], Ag@Au [ 52 , 53 , 54 ], or bimetallic [ 55 , 56 ], SERS probes can be designed. For this, particle Raman reporters can not only be adsorbed onto the nanoparticle surface but also embedded between Au and Ag layers.…”

“…Sánchez-Purrà et al used BPE- and MBA-encoded nanostars as labels for the single test zone simultaneous detection of dengue virus (DENV) and Zika virus (ZIKV). Very recently, Zhang et al [ 54 ] developed a combined multi-tag and multi-T-zone strategy for the simultaneous detection of respiratory viruses. Core shell SERS nanotags encoded with two Raman reporters were chosen as labels and combined on a microarray immobilized on a nitrocellulose membrane for the rapid quantification of the nucleic acids from eleven respiratory viruses on a single strip ( Figure 5 c).…”