key: cord-255075-6azu6k3h authors: Zhuang, Jianjian; Yin, Juxin; Lv, Shaowu; Wang, Ben; Mu, Ying title: Advanced “lab-on-a-chip” to detect viruses – Current challenges and future perspectives date: 2020-05-12 journal: Biosens Bioelectron DOI: 10.1016/j.bios.2020.112291 sha: doc_id: 255075 cord_uid: 6azu6k3h Massive viral outbreaks draw attention to viruses that have not been thoroughly studied or understood. In recent decades, microfluidic chips, known as “lab-on-a-chip”, appears as a promising tool for the detection of viruses. Here, we review the development of microfluidic chips that could be used in response to viral detection, specifically for viruses involved in more recent outbreaks. The advantages as well as the disadvantages of microfluidic systems are discussed and analyzed. We also propose ideas for future development of these microfluidic chips and we expect this advanced technology to be used in the future for viral outbreaks. Viruses infect millions of individuals each year resulting in serious morbidity (Hutchinson 2018 The LOD of this system is 1 copy/μL for SUDV, 100 copies/μL for EBOV, 1000 copies/μL for BDBV 128 and 10 copies/μL for TAFV detected in 50 minutes. 4 minutes) and accurate (as low as 1 copy) detection for EBOV at POC. However, some chips lack 131 sample preparation or require additional instrumentation, which is not suitable in resource-limited 132 settings for POC. Moreover, due to the high contagious rate and characteristics of the multiple EBOV 133 subtypes, low-cost multiplex detection chips should be developed. HIV is a single-stranded RNA virus that results in acquired immunodeficiency syndrome (AIDS) 136 (Watts et al. 2009 ). HIV attacks T lymphocytes and integrates into the chromosomes of its host, which 137 in turn leads to defects in the human immune system causing irreparable damage to the body (Druce et simultaneously. This chip meets serological requirements and detects HIV RNA as low as 10 3 viral 166 particles/ml from saliva or blood. Paper-based microfluidics is also an effective tool for HIV detection. Zhao With the goal of detecting HIV, these studies were mainly performed analyzing CD4 + cells, Currently, the influenza virus is a health concern for "lab on a chip" ( This method utilizes electromagnetically-driven magnetic beads and enzyme-linked immunosorbent 208 (ELISA)-like assays on the platform to detect H1N1 viruses. This system can also reach a LOD of Moreover, rapid and ongoing evolution of influenza viruses will make this even more challenging. In recent years, microfluidics has also shown to be an effective tool for the early diagnosis of (Figure 7 A ) . This strategy used bead beating and heating procedures to 303 obtain stable RNA directly from whole blood, which could be used to detect viruses. In this system, 304 direct buffer was used to extract RNA from whole blood and obtain a LOD of 10 2 PFU/200 μL in less 305 than 1 hour. Yin et al (Yin et al. 2020 ) reported a microfluidic system that integrated RNA extraction 306 and multiplex PCR to detect four DENV serotypes (Figure 7 B ). In this system, chitosan-modified 307 paper chip to extra RNA and on-chip PCR product was detected using a membrane sensor. This system Detection reagents were prestored into the chip and the freeze-dried reagents were kept in 2−8 °C for 6 377 months in the disc chip. Moreover, the separation step of plasma or serum is avoided since it is directly 378 detected from whole blood. This system can detect 10 2 copies/mL HBV DNA in ~48 minutes from 500 379 μL of whole blood. Our group also proposed a digital isothermal chip for the quantitative detection of will give more precise results. In addition, the integration of sample preparation with detection is important especially for 420 nucleic acid-based detection methods (Yin et al. 2019 ). However, very few studies have been able to 421 integrate virus sample preparation into chips. Integrated sample preparation will reduce testing time, improve accuracy and minimize labor. Moreover, sample-in-answer-out is the ideal detection process. Therefore, sample preparation should be considered in a single chip. Whether Lab-on-a-chip is used in 424 the clinic or the home, sample preparation integration is necessary. In addition, affordable and 425 user-friendly qualities should be considered especially for POC in resource-limited settings. Automated and high-throughput microfluidics should also be considered. Therefore, additional sample preparation 427 methods should be tested and integrated into chips that will be used for virus outbreaks. chip is further combined with the "Biological mobile phone", "Mobile detection station", or "Artificial Intelligence"，its potential for virus detection will be extended even further. In the future, microfluidic products that meet the criteria for POC proposed by WHO including (1) 478 being affordable to those at risk of infection, containing (2) high sensitivity, (3) high specificity, (4) 479 user-friendly capabilities, being (5) rapid and robust, (6) equipment free, and (7) Comparison of herpes 603 simplex virus PCR with culture for virus detection in multisource surface swab specimens from 604 neonates Nanoparticle-enhanced electrical detection of Zika virus on paper 607 microchips Improving HIV proteome annotation: new features of BioAfrica HIV Proteomics Resource Multiplexed efficient on-chip sample preparation 613 and sensitive amplification-free detection of Ebola virus Microfluidic 615 System for Detection of Viral RNA in Blood Using a Barcode Fluorescence Reporter Photocleavable Capture Probe Dengue virus: a review on 619 advances in detection and trends -from conventional methods to novel biosensors Differential serum cytokine profiles in patients with chronic hepatitis b, 623 c, and hepatocellular carcinoma The propagation of a Christmas Carol produced by adolescent cancer patients at the Istituto 626 Hands-free smartphone-based diagnostics for simultaneous detection of Zika Dengue at point-of-care Wearable microfluidic diaphragm pressure sensor for health and tactile touch 632 monitoring Microfluidic chips for point-of-care immunodiagnostics Genomic surveillance elucidates Ebola virus origin and 643 transmission during the 2014 outbreak Rapid, low-cost and instrument-free CD4+ cell counting 645 16 for HIV diagnostics in resource-poor settings Prevalence and seasonal distribution of respiratory viruses 647 during the 2014-2015 season in Istanbul The Ebola Epidemic A Global Health Emergency On-chip 651 multiplex electrochemical immunosensor based on disposable 24-site fluidic micro-array screen 652 printing analytical device for multi-component quantitative analysis Point of care diagnostics: status 655 and future Zika virus is a global public health emergency, declares WHO New fronts emerge in the influenza cytokine storm Dengue virus -Mosquito interactions Biology of Zika Virus Infection in Human Skin Cells A multi-virus detectable microfluidic 666 electrochemical immunosensor for simultaneous detection of H1N1, H5N1, and H7N9 virus using ZnO 667 nanorods for sensitivity enhancement Deaths from norovirus 669 among the elderly, England and Wales A comparison study of Zika virus outbreaks 671 in French Polynesia, Colombia and the State of Bahia in Brazil High-throughput and all-solution phase African Swine Fever Virus (ASFV) 674 detection using CRISPR-Cas12a and fluorescence based point-of-care system Traditional and modern cell culture in virus diagnosis. 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A: Ebola virus de (Wit et 1085 al. 2011)；B：Human immunodeficiency virus (Druce et al. 2016)；C：Influenza virus ；E：Dengue virus (Zonetti et al. 2018)；F：SARS-CoV Figure 3：Microfluidic system to detect EBOV. A: Microfluidic chips proposed by Qin et al Adapted from ref. 60 with permission from Nature Publications. C: Paper chip 1114 proposed by Brangel et al. Serum forms complexes between the labeled gold nanoparticles (AuNPs) 1115 and the target analytes. Targeted IgG serum antibodies against single or multiple recombinant Ebola 1116 viral proteins bind to preprinted test lines, forming a visual red-purple line. A control line is used to 1117 validate assay function for the detection of antihuman antibody-gold nanoparticle conjugates. Results 1118 can be obtained in 15 minutes. Adapted from ref. 66 with permission from Nature Publications. D: 1119 Schematic of the microfluidic chip for detection of four EBOV species Figure 4 ：Microfluidic systems for the detection of HIV. A: Schematic and photograph of the manual 1136 magnetophoretic CD4+ isolation chip proposed by Glynn et al. Prior to the test, the chip was perfused 1137 through a degassing process B: Schematic and photograph of the RT-LAMP substrate and smartphone apparatus for HIV 1140 (i) heating stage. The microfluidic chip was put on the stage (ii) copper 1141 base containing mineral oil, (iii) wavelength filters placed in front of the LED and smartphone camera, 1142 (iv) smartphone ,(v) blue LED light source, and (vi) apparatus. Adapted from ref. 79 with permission 1143 from the Elsevier. C：Schematic and photograph of microRAAD for HIV testing When using this system, 1) the paper-based chip should be assembled into plastic housing with a 1145 temperature control circuit, 2) buffer should be added into inlets and sealed with adhesive tape to 1146 minimize evaporation, 3) the chip should be connected with a phone to heat, 4) one should wait 90 1147 minutes for automated fluid delivery and sample incubation in μPAD D: Schematic of a 1149 paper-based chip for the detection of HIV developed by Li et al. Image shows the components (left) 1150 and the assembly (right) of origami chip This is a magnetism and size mediated platform. Different 1172 influenza subtypes could be simultaneously separated and detected depending on the different-sizes 1173 magnetic beads. Adapted from ref. 106 with permission from the Elsevier. D: Photograph microfluidic 1174 chip for the detection of 12 influenza subtypes (Shen et al 2019). Arrayed reaction chambers contain 1175 primer sets for amplifying specific regions of the HA and NA genes such that the RT-PCR-derived 1176 signal output could be used for viral subtyping Figure 6： Microfluidic systems for the detection of ZIKV. A: Workflow of the portable cup device 1188 based on the microfluidic system developed by Song et al. (a) Schematic of saliva sample preparation Saliva samples are collected in a saliva collection tube and then lysed in Qiagen binding/lysis (AVL) The lysed sample is filtered through the isolation membrane of a microfluidic cassette for 1191 nucleic acid extraction. (c) Enlarged view of the chemically-heated cup. The cup consists of a thermos 1192 cup body, a 3Dprinted cup lid, a chip holder, PCM material, heat sink and single-use Mg-Fe alloy pack 1193 heat source. (d) A photograph of the chemically-heated cup for point of care molecular diagnostics of 1194 ZIKV B: Schematic of the wearable microfluidic sensor for detection of ZIKV nucleic acids The detection method was based on the RPA. The reaction can start on this wearable microfluidic 1197 system effectively by human epidermal heat that was close to the optimal temperature of RPA. Adapted 1198 from ref. 123 with permission from the Elsevier The performance of microfluidic systems in virus detection are discussed and analyzed The challenges of microfluidic systems with regard to sample preparation, throughput, and multiplexing are highlighted Ideas for future development of microfluidic systems were proposed. ☒ The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.☐The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: