ABSTRACT
Since the outbreak of coronavirus disease 2019 (COVID-19), efficient real-time monitoring has become one of the challenges faced in SARS-CoV-2 virus detection. A compact all-fiber Mach-Zehnder interferometer optofluidic sensor based on a hollow eccentric core fiber (HECF) for the detection and real-time monitoring of SARS-CoV-2 spike glycoprotein (SARS-CoV-2 S2) is proposed, analyzed and demonstrated. The sensor is comprised of fusion splicing single mode fiber (SMF), hollow core fiber (HCF) and HECF. After the incident light passes through the HCF from the SMF, it uniformly enters the air hole and the suspended micrometer-scale fiber core of the HECF to form a compact all-fiber Mach-Zehnder interferometer (MZI). HECF is side polished to remove part of the cladding that the suspended fiber core can contact the external environment. Subsequently, the mouse anti SARS-CoV-2 S2 antibody is fixed on the surface of the suspended-core for the sake of achieving high sensitivity and specific sensing of SARS-CoV-2 S2. The limit of detection (LOD) of the sensor is 26.8 pM. The proposed sensor has high sensitivity, satisfactory selectivity, and can be fabricated at low cost making it highly suitable for point-of-care testing and high-throughput detection of early stage of COVID-19 infection.
ABSTRACT
Herein, a pipette-tip-enabled digital nucleic acid analyzer for high-performance COVID-19 testing is demonstrated. This is achieved by digital loop-mediated isothermal amplification (digital LAMP or dLAMP) using common laboratory equipment and materials. It is shown that simply fixing a glass capillary inside conventional pipette tips enables the generation of monodisperse, water-in-oil microdroplets with benchtop centrifugation. It is shown that using LAMP, the ORF1a/b gene, a standard test region for COVID-19 screening, can be amplified without a thermal cycler. The amplification allows counting of fluorescent microdroplets so that Poisson analysis can be performed to allow quantification with a limit of detection that is 1 order of magnitude better than those of nondigital techniques and comparable to those of commercial dLAMP platforms. It is envisioned that this work will inspire studies on ultrasensitive digital nucleic acid analyzers demanding both sensitivity and accessibility, which is pivotal to their large-scale applications.
Subject(s)
COVID-19 Testing/methods , COVID-19 , Nucleic Acid Amplification Techniques , COVID-19/diagnosis , Humans , Molecular Diagnostic TechniquesABSTRACT
In this work, it is shown that surface-enhanced Raman scattering (SERS) measurements can be performed using liquid platforms to perform bioanalysis at sub-pM concentrations. Using magnetic enrichment with gold-coated magnetic nanoparticles, the high sensitivity was verified with nucleic acid and protein targets. The former was performed with a DNA fragment associated with the bacteria Staphylococcus aureus, and the latter using IgG antibody, a biomarker for COVID-19 screening. It is anticipated that this work will inspire studies on ultrasensitive SERS analyzers suitable for large-scale applications, which is particularly important for in vitro diagnostics and environmental studies.