Method For Detection Of Virus Research Articles

Quick and ultra-sensitive digital assay of influenza virus using sub-picoliter microwells

Quick and sensitive virus detection methods have been developed using sub-picoliter microwells. One method uses an aggregation-induced emission (AIE) reagent, and the other uses an enzymatic reaction supported with immunomagnetic beads at a high concentration of 108 particles/mL. Examination of influenza A virus detection using the AIE reagent exhibited a detection limit of 3 × 105 copies/mL, which was achievable within 1 min of the total measurement time. The high-concentration immunomagnetic beads method exhibited a detection limit as low as 1 × 102 copies/mL. The developed methods are effective and practical tools for ultrafast and ultrasensitive virus detection.

Evaluation of conventional and point-of-care real-time RT-PCR tests for the detection of SARS-CoV-2 through a pooled testing strategy.

BackgroundThe rapid identification and isolation of individuals infected with SARS‐CoV‐2 are fundamental countermeasures for the efficient control of the COVID‐19 pandemic, which has affected millions of people around the world. Real‐time RT‐PCR is one of the most commonly applied reference methods for virus detection, and the use of pooled testing has been proposed as an effective way to increase the throughput of routine diagnostic tests. However, the clinical applicability of different types of real‐time RT‐PCR tests in a given group size remains inconclusive due to inconsistent regional disease prevalence and test demands.MethodsIn this study, the performance of one dual‐target conventional and two point‐of‐care real‐time RT‐PCR tests in a 5‐specimen pooled testing strategy for the detection of SARS‐COV‐2 was evaluated.ResultsWe demonstrated the proof of concept that all of these real‐time RT‐PCR tests could feasibly detect SARS‐CoV‐2 from nasopharyngeal and oropharyngeal specimens that contain viral RNA loads in the range of 3.48 × 105 to 3.42 × 102 copies/ml through pooled testing in a group size of 5 with overall positive percent agreement (pooling vs. individual testing) ranging from 100% to 93.75%. Furthermore, the two POC real‐time RT‐PCR tests exhibited comparable sensitivity to that of the dual‐target conventional one when clinical specimens were tested individually.ConclusionOur findings support the feasibility of using real‐time RT‐PCR tests developed as a variety of platforms in routine laboratory detection of suspected COVID‐19 cases through a pooled testing strategy that is beneficial to increasing the daily diagnostic capacity.

Respiratory virus detection in the upper respiratory tract of asymptomatic, community-dwelling older people

BackgroundThe prevalence of virus positivity in the upper respiratory tract of asymptomatic community-dwelling older people remains elusive. Our objective was to investigate the prevalence of respiratory virus PCR positivity in asymptomatic community-dwelling older people using saliva samples and nasopharyngeal and oropharyngeal swabs.MethodsWe analyzed 504 community-dwelling adults aged ≥ 65 years who were ambulatory and enrolled in a cross-sectional study conducted from February to December 2018 in Nagasaki city, Japan. Fourteen respiratory viruses were identified in saliva, nasopharyngeal and oropharyngeal samples using multiplex PCR assays.ResultsThe prevalences of PCR positivity for rhinovirus, influenza A, enterovirus and any respiratory virus were 12.9% (95% CI: 10.1–16.1%), 7.1% (95% CI: 5.1–9.8%), 6.9% (95% CI: 4.9–9.5%) and 25.2% (95% CI: 21.5–29.2%), respectively. Rhinovirus was detected in 21.5% of subjects, influenza A in 38.9% of subjects, enterovirus in 51.4% of subjects and any virus in 32.3% of subjects using only saliva sampling.ConclusionsThe prevalences of several respiratory viruses were higher than the percentages reported previously in pharyngeal samples from younger adults. Saliva sampling is a potentially useful method for respiratory virus detection in asymptomatic populations.

Optimizing sample collection methods for detection of respiratory viruses in poultry housing environments.

Viral respiratory diseases, such as avian influenza, Newcastle disease, infectious bronchitis and infectious laryngotracheitis, have considerable negative economic implications for poultry. Ensuring the virus-free status of premises by environmental sampling after cleaning and disinfection is essential for lifting a quarantine and/or safely restocking the premises following an outbreak. The objectives of this study were to identify optimal sample collection devices and to determine the locations in poultry housing which are best for poultry respiratory virus sample collection. Chickens exposed to infectious bronchitis virus, which was used as a representative virus for enveloped poultry respiratory viruses, were housed in floor-pens in either a curtain-sided wood framed house or a cement block house. Foam swabs, cellulose sponges, polyester swabs, dry cotton gauze and pre-moistened cotton gauze were evaluated for comparative efficiency in recovering viral RNA. Cotton gauze pre-moistened with the viral transport media had the highest sensitivity among the devices (wood-framed house: 78% positive, geometric mean titre [GMT] of 2.6 log10 50% egg infectious doses [EID50 ] equivalents/ml; cement block houses: 55% positive, GMT of 1.7 log10 EID50 equivalents/ml). Targeting virus deposition sites is also crucial for efficient virus elimination procedures and subsequent testing; therefore, 10 locations within the houses were compared for virus detection. In both housing types, the highest viral RNA loads were recovered from the tops of drinker lines within the pen. Places the chickens could contact directly (e.g., feeder rim) or were contacted by caretaker feet (hallway floor) also yielded higher levels of viral RNA more consistently. These results will facilitate the establishment of efficient environmental sampling procedures for respiratory viruses of poultry.

Virus Detection using Second Harmonics of Magnetic Nanoparticles

Virus detection methods based on the nonlinear magnetic response of magnetic nanoparticles have been investigated, and magnetic detection methods using the third harmonic have been widely applied on account of their high sensitivity, short measurement‐time, and low cost. In this letter, we propose a virus detection method using the second harmonics to improve the signal intensity. We find that the signal to noise ratio of the second harmonic is approximately three times higher than that of the third harmonic. A comparison of the ratio of the second harmonic to the fourth harmonic (R24) and the ratio of the third harmonic to the fifth harmonic (R35) shows that R24 is more sensitive to identifying changes in virus concentration. Our proposed method has the potential to be utilized for rapid screening on virus detection. © 2022 Institute of Electrical Engineers of Japan. Published by Wiley Periodicals LLC.

Virus Detection: From State-of-the-Art Laboratories to Smartphone-Based Point-of-Care Testing.

Infectious virus outbreaks pose a significant challenge to public healthcare systems. Early and accurate virus diagnosis is critical to prevent the spread of the virus, especially when no specific vaccine or effective medicine is available. In clinics, the most commonly used viral detection methods are molecular techniques that involve the measurement of nucleic acids or proteins biomarkers. However, most clinic‐based methods require complex infrastructure and expensive equipment, which are not suitable for low‐resource settings. Over the past years, smartphone‐based point‐of‐care testing (POCT) has rapidly emerged as a potential alternative to laboratory‐based clinical diagnosis. This review summarizes the latest development of virus detection. First, laboratory‐based and POCT‐based viral diagnostic techniques are compared, both of which rely on immunosensing and nucleic acid detection. Then, various smartphone‐based POCT diagnostic techniques, including optical biosensors, electrochemical biosensors, and other types of biosensors are discussed. Moreover, this review covers the development of smartphone‐based POCT diagnostics for various viruses including COVID‐19, Ebola, influenza, Zika, HIV, et al. Finally, the prospects and challenges of smartphone‐based POCT diagnostics are discussed. It is believed that this review will aid researchers better understand the current challenges and prospects for achieving the ultimate goal of containing disease‐causing viruses worldwide.

Virus detection methods for different kinds of food and water samples – The importance of molecular techniques

The microbiological safety of food and the environment in which we live is currently an intensely discussed topic. Increasing production and the demand-driven global market exert pressure on ensuring sufficient high-quality food and safe drinking water. Compared to the past, increased attention in this area is now paid to important viral agents associated with food/water contaminations in both intensive research and routine diagnostics. This interest in viral agents has also increased in recent years due to the ongoing global pandemic caused by the novel coronavirus. Food- and water-borne viruses usually cause only mild and short-term diseases. The most common is gastroenteritis manifested by fever, vomiting and watery diarrhoea. However, in addition to mild febrile illness, these agents can also cause more serious conditions – respiratory infections, hepatitis, conjunctivitis, aseptic meningitis, myocarditis, encephalitis and paralysis. Globally, these diseases have significant economic impacts and are still among the leading causes of death in developing countries. This manuscript provides an overview of food- and water-borne viruses and technologies developed and currently used for their identification as causative agents. Methods for the detection of these pathogens represent an important tool for the assessment and mitigation of potential risks associated with the contamination of food and water resources. There is currently a wide range of possible approaches. Their use is differently targeted and their sensitivity, effectiveness and specificity also vary. In the case of a specific application, it is therefore necessary to choose the appropriate method, optimize it, and then verify its applicability and limits. The chosen method should be sufficiently robust, sensitive, specific and, if possible, also time and labor saving.

Split T7 promoter-based isothermal transcription amplification for one-step fluorescence detection of SARS-CoV-2 and emerging variants

The negative global impact of the coronavirus disease pandemic has highlighted the crucial need for a rapid and convenient method of viral RNA detection. In this study, we report a novel method, termed as the split T7 promoter-based isothermal transcription amplification with light-up RNA aptamer (STAR), for one-pot detection of viral RNA. STAR uses a split T7 promoter that is applied to a three-way junction to mediate the selective transcription by the T7 RNA polymerase in the presence of target RNA. In addition, a light-up RNA aptamer is used for signal amplification. STAR can detect viral RNA in less than 30 min with high specificity and sensitivity. By testing of 60 nasopharyngeal SARS-CoV-2 samples, the STAR assay demonstrates an excellent sensitivity and specificity of 96.7% and 100%, respectively. Moreover, we provide experimental evidence of the broad applicability of this assay through the multiplex detection of SARS-CoV-2 variants (D614G mutation) and direct detection of bacterial 16S rRNA.

Saliva collection via capillary method may underestimate arboviral transmission by mosquitoes

BackgroundArthropod-borne viruses (arboviruses) impose a major health and economic burden on human populations globally, with mosquitoes serving as important vectors. Measuring the ability of a mosquito population to transmit an arbovirus is important in terms of evaluating its public health risk. In the laboratory, a variety of methods are used to estimate arboviral transmission by mosquitoes, including indirect methods involving viral detection from mosquito saliva collected by forced salivation. The accuracy of indirect methods to estimate arbovirus transmission to live animal hosts has not been fully evaluated.MethodsWe compared three commonly used proxies of arboviral transmission, namely, the presence of virus in mosquito legs, in salivary glands (SG) and in saliva collected in capillary tubes using forced salivation, with direct transmission estimates from mosquitoes to suckling mice. We analyzed five vector-virus combinations, including Aedes aegypti infected with chikungunya virus, West Nile virus and Zika virus; Culex quinquefasciatus infected with West Nile virus; and Aedes triseriatus infected with La Crosse virus.ResultsComparatively, the methods of detecting virus infection in mosquito legs and in SG were equally accurate in predicting transmission. Overall, the presence of virus in mosquito legs was a more accurate predictor of transmission than the commonly implemented viral detection method using forced salivation into a capillary tube, and was subject to less technical variation.ConclusionsThese results suggest that, in general, forced salivation methods tend to underestimate virus transmission, and they provide confidence in the use of mosquito leg screens to evaluate the transmission potential of a mosquito population.Graphical

RPA-Cas12aDS: A visual and fast molecular diagnostics platform based on RPA-CRISPR-Cas12a method for infectious bursal disease virus detection

Infectious bursal disease (IBD), a major disease of birds, is caused by infectious bursal disease virus (IBDV). The disease can lead to immunosuppression, resulting in huge economic losses in the poultry industry. A specific, rapid, and simple detection method is important for the early diagnosis and prevention and control of IBDV. In this study, we established a naked-eye visual IBDV detection method, named “RPA-Cas12aDS”, by combining recombinase polymerase amplification (RPA) with CRISPR-Cas12a-based nucleic acid detection. The detection process can be accomplished in 50 min, and uncapping contamination can be avoided. The detection results can be observed under blue or UV light. We used the RPA-Cas12aDS method to detect IBDV in bursa of Fabricius tissue samples of chickens, and the results were consistent with those obtained using commercial RT-PCR kits. This method presents great potential for visual, rapid, and point-of-care molecular diagnostics of IBDV in poultry.

A Multi-Laboratory Comparison of Methods for Detection and Quantification of African Swine Fever Virus.

African swine fever is a viral disease of the family Suidae. Methods to detect and quantify African swine fever virus (ASFV) include qPCR and virus infectivity assays. Individual laboratories often use in-house procedures for these assays, which can hamper the comparison of results. The objective of this study was to estimate the probability of ASFV detection using these assays, and to determine the inter-test correlations between results. This was achieved by testing a panel of 80 samples at three reference laboratories. Samples were analysed using nucleic acid extraction and qPCR, as well as virus infectivity assays. For qPCR, a very high probability (ranging from 0.96 to 1.0) of detecting ASFV DNA was observed for all tested systems. For virus infectivity assays in cells, the probability of detecting infectious ASFV varied from 0.68 to 0.90 and was highest using pulmonary alveolar macrophages, followed by MARC145 cells, peripheral blood monocytes, and finally wild boar lung cells. Intraclass correlation coefficient estimates of 0.97 (0.96–0.98) between qPCR methods, 0.80 (0.74–0.85) to 0.94 (0.92–0.96) between virus infectivity assays, and 0.77 (0.68–0.83) to 0.95 (0.93–0.96) between qPCR methods and virus infectivity assays were obtained. These findings show that qPCR gives the highest probability for the detection of ASFV.

Toward Mail-in-Sensors for SARS-CoV-2 Detection: Interfacing Gel Switch Resonators with Cell-Free Toehold Switches

The COVID-19 pandemic has emphasized the importance of widespread testing to control the spread of infectious diseases. The rapid development, scale-up, and deployment of viral and antibody detection methods since the beginning of the pandemic have greatly increased testing capacity. Desirable attributes of detection methods are low product costs, self-administered protocols, and the ability to be mailed in sealed envelopes for the safe analysis and subsequent logging to public health databases. Herein, such a platform is demonstrated with a screen-printed, inductor-capacitor (LC) resonator as a transducer and a toehold switch coupled with cell-free expression as the biological selective recognition element. In the presence of the N-gene from SARS-CoV-2, the toehold switch relaxes, protease enzyme is expressed, and it degrades a gelatin switch that ultimately shifts the resonant frequency of the planar resonant sensor. The gelatin switch resonator (GSR) can be analyzed through a sealed envelope allowing for assessment without the need for careful sample handling with personal protective equipment or the need for workup with other reagents. The toehold switch used in this sensor demonstrated selectivity to SARS-CoV-2 virus over three seasonal coronaviruses and SARS-CoV-1, with a limit of detection of 100 copies/μL. The functionality of the platform and assessment in a sealed envelope with an automated scanner is shown with overnight shipment, and further improvements are discussed to increase signal stability and further simplify user protocols toward a mail-in platform.

Numerical approach to design the graphene-based multilayered surface plasmon resonance biosensor for the rapid detection of the novel coronavirus

In this article, a graphene-based multilayered surface plasmon resonance (SPR) biosensor of (BK7/WS2/Au/BaTiO3/Graphene) is proposed for the rapid detection of the novel coronavirus (COVID-19). The proposed SPR biosensor is designed based on the angular interrogation attenuated total reflection (ATR) method for rapid detection of the COVID-19 virus. The sensor’s surface plasmon polaritons (SPPs) and the sensing region refractive index (RI) are changed, owing to the interaction of various concentrated ligand-analytes. The specific ligand is mechanized with the proposed sensor surface and the target analyte that has flowed onto the sensing surface. The proposed sensor is capable of detecting the COVID-19 virus rapidly in two different ligand-analytes environments, such as: (i) the virus spike receptor-binding domain (RBD) as an analyte and monoclonal antibodies (mAbs) as a probe ligand, and (ii) the monoclonal antibodies (IgG or IgM) as an analyte and the virus spike RBD as a probe ligand. Due to the binding of the target ligand-analytes, the concentration level of the sensing region is incremented. As the increment in the concentration level, the RI of the sensing medium increases, therefore the change in RI causes the shift in the SPR angle resulting in the output reflectance intensity. The performance of the multilayered SPR sensor is analyzed numerically using the finite element method (FEM) method. Numerically, the proposed sensor provides the maximum angular shift sensitivity at 230.77 deg/refractive index unit (RIU), detection accuracy (DA) at 0.161 deg−1, and the figure of merits (FOM) is at 37.22 RIU−1. In addition, with each additional graphene layer number (L), the proposed sensor exhibits the angular shift sensitivity increment (1 + 0.7L) times. The novelty of the proposed multilayer (BK7/WS2/Au/BaTiO3/Graphene) sensor is highly angular sensitivity, and capable of detecting the COVID-19 virus rapidly without a false-positive report.

First case of SARS-CoV-2 RNA detection in municipal solid waste leachate from Brazil

This work presents the first case of SARS-CoV-2 RNA detection in leachate collected from a transfer station in the city of São Paulo, Brazil. After calibration of the viral detection method already used for wastewater samples with a pilot leachate sample and virus fragments in laboratory, twelve polyethylene glycol concentrated leachates samples were tested by RT-qPCR. The results confirmed the presence of N1 gene in 9 of the 12 analyzed samples between epidemiological weeks 33 and 38 of the year 2021 (08/15/2021 to 09/19/2021). The occurrence of the N2 gene was only observed in 5 of the 12 samples. The concentration values for N1 and N2 genes varied between 3.1 and 4.6 log10.GC·L−1, which are values close to those measured in sanitary wastewater. This method showed to be a promising procedure to verify the presence of viral RNA in municipal solid waste leachate, being especially useful where there is no treatment system and sanitation infrastructure, which makes the conventional wastewater surveillance unfeasible.

Method for tick-borne encephalitis virus detection in raw milk products

The transmission of tick-borne encephalitis virus (TBEV) through food is rare, but can occur through the consumption of raw milk products from animals infected by tick bites. In 2020, France faced a TBEV outbreak linked to the consumption of unpasteurized goat cheese. The aim of this study was to develop and characterize a molecular method for the detection of TBEV in raw milk products based on the recent international standard PR ISO/DIS 16140-4. The TBEV recovery rates varied with the inoculation level and settings. The LOD50 and LOD95 of TBEV were 6.40 × 103 genome copies per g or per mL and 2.84 × 104 genome copies per g or per mL, respectively. The percentages of RT-qPCR inhibitions were lower than 75% and the murine norovirus (MNV-1), used as process control, was detected in all samples with a recovery rate higher than 1%, as recommended in ISO 15216. We conclude that the described method is appropriate to detect TBEV in raw milk products for routine diagnosis, and to assess potential health risks.

Optimization of Silicon Nitride Waveguide Platform for On-Chip Virus Detection.

This work presents a rigorous and generic sensitivity analysis of silicon nitride on silicon dioxide strip waveguide for virus detection. In general, by functionalizing the waveguide surface with a specific antibodies layer, we make the optical sensor sensitive only to a particular virus. Unlike conventional virus detection methods such as polymerase chain reaction (PCR), integrated refractive index (RI) optical sensors offer cheap and mass-scale fabrication of compact devices for fast and straightforward detection with high sensitivity and selectivity. Our numerical analysis includes a wide range of wavelengths from visible to mid-infrared. We determined the strip waveguide’s single-mode dimensions and the optimum dimensions that maximize the sensitivity to the virus layer attached to its surface at each wavelength using finite difference eigenmode (FDE) solver. We also compared the strip waveguide with the widely used slot waveguide. Our theoretical study shows that silicon nitride strip waveguide working at lower wavelengths is the optimum choice for virus detection as it maximizes both the waveguide sensitivity (Swg) and the figure of merit (FOM) of the sensor. The optimized waveguides are well suited for a range of viruses with different sizes and refractive indices. Balanced Mach–Zehnder interferometer (MZI) sensors were designed using FDE solver and photonic circuit simulator at different wavelengths. The designed sensors show high FOM at λ = 450 nm ranging from 500 RIU−1 up to 1231 RIU−1 with LMZI = 500 µm. Different MZI configurations were also studied and compared. Finally, edge coupling from the fiber to the sensor was designed, showing insertion loss (IL) at λ = 450 nm of 4.1 dB for the design with FOM = 500 RIU−1. The obtained coupling efficiencies are higher than recently proposed fiber couplers.

Protein Palmitoylation Modification During Viral Infection and Detection Methods of Palmitoylated Proteins.

Protein palmitoylation—a lipid modification in which one or more cysteine thiols on a substrate protein are modified to form a thioester with a palmitoyl group—is a significant post-translational biological process. This process regulates the trafficking, subcellular localization, and stability of different proteins in cells. Since palmitoylation participates in various biological processes, it is related to the occurrence and development of multiple diseases. It has been well evidenced that the proteins whose functions are palmitoylation-dependent or directly involved in key proteins’ palmitoylation/depalmitoylation cycle may be a potential source of novel therapeutic drugs for the related diseases. Many researchers have reported palmitoylation of proteins, which are crucial for host-virus interactions during viral infection. Quite a few explorations have focused on figuring out whether targeting the acylation of viral or host proteins might be a strategy to combat viral diseases. All these remarkable achievements in protein palmitoylation have been made to technological advances. This paper gives an overview of protein palmitoylation modification during viral infection and the methods for palmitoylated protein detection. Future challenges and potential developments are proposed.

Detection of NS1 protein from dengue virus in excreta and homogenates of wild-caught Aedes aegypti mosquitoes using monoclonal antibodies.

Dengue fever is one of the most devastating infectious diseases worldwide. Development of methods for dengue virus (DENV) detection in mosquitoes to assess prevalence as a preliminary screen for entomological surveillance in endemic regions of DENV will certainly contribute to the control of the disease. A monoclonal antibody against the NS1 (nonstructural protein 1) viral protein was generated using recombinant NS1 protein and used to detect and analyze DENV in both excreta and total homogenates from Aedes aegypti mosquitoes. Results demonstrated expression of NS1 in excreta of DENV laboratory-infected mosquitoes and homogenates from field mosquitoes infected with DENV. The immunodetection method reported here represents a first-line strategy for assessing the prevalence of DENV in mosquitoes, for entomological surveillance in endemic regions of dengue. Detection of DENV prevalence in field mosquitoes could have an impact on vector surveillance measures to interrupt dengue transmission.

Electrochemical biosensor for nucleic acid amplification-free and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA via CRISPR/Cas13a trans-cleavage reaction

The outbreak of the COVID-19 pandemic has led to millions of fatalities worldwide. For preventing epidemic transmission, rapid and accurate virus detection methods to early identify infected people are urgently needed in the current situation. Therefore, an electrochemical biosensor based on the trans-cleavage activity of CRISPR/Cas13a was developed in this study for rapid, sensitive, and nucleic-acid-amplification-free detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Herein, a redox probe conjugated with ssRNA is immobilized on the electrode surface modified with a nanocomposite (NC) and gold nanoflower (AuNF) for enhancing the sensing performance. The SARS-CoV-2 RNA is captured by the Cas13a–crRNA complex, which triggers the RNase function of Cas13a. The enzymatically activated Cas13a–crRNA complex is subsequently introduced to the reRNA-conjugated electrochemical sensor, and consequently cleaves the reRNA. A change in current occurs due to the release of the redox molecule labeled on the reRNA, which is trans-cleaved from the Cas13a–crRNA complex. The biosensor can detect as low as 4.4 × 10−2 fg/mL and 8.1 × 10−2 fg/mL of ORF and S genes, respectively, over a wide dynamic range (1.0 × 10−1 to 1.0 × 105 fg/mL). Moreover, the biosensor was evaluated by measuring SARS-CoV-2 RNA spiked in artificial saliva. The recovery of the developed sensor was found to be in an agreeable range of 96.54–101.21%. The designed biosensor lays the groundwork for pre-amplification-free detection of ultra-low concentrations of SARS-CoV-2 RNA and on-site and rapid diagnostic testing for COVID-19.

Biosensor detection of airborne respiratory viruses such as SARS-CoV-2.

Airborne SARS-CoV-2 transmission represents a significant route for possible human infection that is not yet fully understood. Viruses in droplets and aerosols are difficult to detect because they are typically present in low amounts. In addition, the current techniques used, such as RT-PCR and virus culturing, require large amounts of time to get results. Biosensor technology can provide rapid, handheld, and point-of-care systems that can identify virus presence quickly and accurately. This paper reviews the background of airborne virus transmission and the characteristics of SARS-CoV-2, its relative risk for transmission even at distances greater than the currently suggested 6 feet (or 2 m) physical distancing. Publications on biosensor technology that may be applied to the detection of airborne SARS-CoV-2 and other respiratory viruses are also summarized. Based on the current research we believe that there is a pressing need for continued research into handheld and rapid methods for sensitive collection and detection of airborne viruses. We propose a paper-based microfluidic chip and immunofluorescence assay as one method that could be investigated as a low-cost and portable option.