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1.
Biosens Bioelectron ; 222: 114987, 2023 Feb 15.
Article in English | MEDLINE | ID: covidwho-2235818

ABSTRACT

Accurate COVID-19 screening via molecular technologies is still hampered by bulky instrumentation, complicated procedure, high cost, lengthy testing time, and the need for specialized personnel. Herein, we develop point-of-care upconversion luminescence diagnostics (PULD), and a streamlined smartphone-based portable platform facilitated by a ready-to-use assay for rapid SARS-CoV-2 nucleocapsid (N) gene testing. With the complementary oligo-modified upconversion nanoprobes and gold nanoprobes specifically hybridized with the target N gene, the luminescence resonance energy transfer effect leads to a quenching of fluorescence intensity that can be detected by the easy-to-use diagnostic system. A remarkable detection limit of 11.46 fM is achieved in this diagnostic platform without the need of target amplification, demonstrating high sensitivity and signal-to-noise ratio of the assay. The capability of the developed PULD is further assessed by probing 9 RT-qPCR-validated SARS-CoV-2 variant clinical samples (B.1.1.529/Omicron) within 20 min, producing reliable diagnostic results consistent with those obtained from a standard fluorescence spectrometer. Importantly, PULD is capable of identifying the positive COVID-19 samples with superior sensitivity and specificity, making it a promising front-line tool for rapid, high-throughput screening and infection control of COVID-19 or other infectious diseases.


Subject(s)
Biosensing Techniques , COVID-19 , Humans , COVID-19/diagnosis , SARS-CoV-2/genetics , Point-of-Care Systems , RNA, Viral/genetics , Luminescence , Smartphone , Biosensing Techniques/methods , Sensitivity and Specificity
2.
J Med Virol ; 95(1): e28397, 2023 01.
Article in English | MEDLINE | ID: covidwho-2157850

ABSTRACT

The information provided by SARS-CoV-2 spike (S)-targeting immunoassays can be instrumental in clinical-decision making. We compared the performance of the Elecsys® Anti-SARS-CoV-2 S assay (Roche Diagnostics) and the LIAISON® SARS-CoV-2 TrimericS IgG assay (DiaSorin) using a total of 1176 sera from 797 individuals, of which 286 were from vaccinated-SARS-CoV-2/experienced (Vac-Ex), 581 from vaccinated/naïve (Vac-N), 147 from unvaccinated/experienced (Unvac-Ex), and 162 from unvaccinated/naïve (Unvac-N) individuals. The Roche assay returned a higher number of positive results (907 vs. 790; p = 0.45; overall sensitivity: 89.3% vs. 77.6%). The concordance between results provided by the two immunoassays was higher for sera from Vac-N (Ï°: 0.58; interquartile ranges [IQR]: 0.50-0.65) than for sera from Vac-Ex (Ï°: 0.19; IQR: -0.14 to 0.52) or Unvac-Ex (Ï°: 0.18; IQR: 0.06-0.30). Discordant results occurred more frequently among sera from Unvac-Ex (34.7%) followed by Vac-N (14.6%) and Vac-Ex (2.7%). Antibody levels quantified by both immunoassays were not significantly different when <250 (p = 0.87) or <1000 BAU/ml (p = 0.13); in contrast, for sera ≥1000 BAU/ml, the Roche assay returned significantly higher values than the DiaSorin assay (p < 0.008). Neutralizing antibody titers (NtAb) were measured in 127 sera from Vac-Ex or Vac-N using a S-pseudotyped virus neutralization assay of Wuhan-Hu-1, Omicron BA.1, and Omicron BA.2. The correlation between antibody levels and NtAb titers was higher for sera from Vac-N than those from Vac-Ex, irrespective of the (sub)variant considered. In conclusion, neither qualitative nor quantitative results returned by both immunoassays are interchangeable. The performance of both assays was found to be greatly influenced by the vaccination and SARS-CoV-2 infection status of individuals.


Subject(s)
COVID-19 Vaccines , COVID-19 , Humans , Luminescence , COVID-19/diagnosis , SARS-CoV-2 , Vaccination , Antibodies, Viral , Immunoglobulin G , Antibodies, Neutralizing , Immunoassay
3.
Anal Chem ; 94(48): 16967-16974, 2022 Dec 06.
Article in English | MEDLINE | ID: covidwho-2133136

ABSTRACT

Capable of precise simultaneous multitarget identifications within a minimized sample, optical multiplexing is vital for accurate diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) while remaining spectral crowding and background interfering. In merits of an autofluorescence-free background and high-capability throughput, a persistent luminescence (PersL) lifetime/color binary encoding strategy was herein proposed for SARS-CoV-2 diagnosis. Based on luminescence resonance energy transfer processes, the intense lifetimes and representative emissions of PersL nanoplatforms were rationally manipulated to create a temporal coding dimension within a wide seconds-to-minutes range through three individual channels. Particularly, at least four populations of barcoding in a certain channel were successfully decoded by a purpose-built time-resolved PersL technology. As a proof-of-concept, functionalized PersL nanoplatforms were further well developed for the simultaneous quantification of five-plex SARS-CoV-2 biomarkers with limits of detection in the subnanomolar range. Remarkably, PersL nanoplatforms enabled a highly differentiable discrimination of multitargets at various concentrations of ultralow background and high-fidelity resolutions, thereby advancing a powerful tool for optical multiplexing in biomedical applications.


Subject(s)
COVID-19 , Luminescence , Humans , SARS-CoV-2 , COVID-19 Testing , COVID-19/diagnosis , Fluorescence Resonance Energy Transfer
4.
Anal Chem ; 94(33): 11591-11599, 2022 08 23.
Article in English | MEDLINE | ID: covidwho-1984346

ABSTRACT

The COVID-19 pandemic has led to a global crisis with devastating effects on public healthcare and the economy. Sensitive detection of SARS-CoV-2 is the key to diagnose and control its spread. The spike (S) protein is an abundant viral transmembrane protein and a suitable target protein for the selective recognition of SARS-CoV-2. Here, we report that with bovine serum albumin prescreening, a specific phage peptide targeting SARS-CoV-2 S1 protein was biopanned with the pIII phage display library. The identified phage #2 expressing the peptide (amino acid sequence: NFWISPKLAFAL) shows high affinity to the target with a dissociation constant of 3.45 ± 0.58 nM. Furthermore, the identified peptide shows good specificity with a binding site at the N-terminal domain of the S1 subunit through a hydrogen bond network and hydrophobic interaction, supported by molecular docking. Then, a sandwiched phage-based enzyme-linked chemiluminescence immunoassay (ELCLIA) was established by using phage #2 as a bifunctional probe capable of SARS-CoV-2 S1 antigen recognition and signal amplification. After optimizing the conditions, the proposed phage ELCLIA exhibited good sensitivity, and as low as 78 pg/mL SARS-CoV-2 S1 could be detected. This method can be applied to detect as low as 60 transducing units (TU)/mL SARS-CoV-2 pseudovirus in 50% saliva. Therefore, specific phage peptides have good prospects as powerful biological recognition probes for immunoassay detection and biomedical applications.


Subject(s)
Bacteriophages , COVID-19 , COVID-19/diagnosis , Humans , Immunoassay , Luminescence , Molecular Docking Simulation , Pandemics , Peptides , SARS-CoV-2
5.
Virus Res ; 319: 198852, 2022 Oct 02.
Article in English | MEDLINE | ID: covidwho-1984220

ABSTRACT

BACKGROUND: Neutralizing antibodies (NAbs) against SARS-CoV-2 infection have a pivotal role in protective immune response; however, their measurement requires specialized facilities. We evaluated the degree of correlation between NAbs and anti-SARS-CoV-2 IgG/total Ig antibodies detected by chemiluminescent immunoassay in asymptomatic and previously symptomatic SARS-CoV-2 patients. METHODS: A total of 1241 participants (previously symptomatic patients and asymptomatic individuals), who were screened for SARS-CoV-2 infection by RT-PCR or serology, were enrolled in our study. Sera were analyzed for the presence of anti-spike-1(S1)-SARS-CoV-2 IgG/total Ig antibodies, using Ortho Clinical Diagnostics, USA. A signal/cut-off value (S/CO) ≥ 1 was considered reactive. NAbs were measured in 103 random samples from groups using microneutralization assay, with titer ≥ 1:10 being considered positive. RESULTS: Asymptomatic (n = 229) and 261 previously symptomatic individuals with positive serology and negative RT-PCR were finally included. Significant higher anti-S1-IgG titers were seen in asymptomatic individuals (P < 0.0001). Conversely, anti-S1-total Ig titers were significantly higher in previously symptomatic (P < 0.0001). NAbs were detected in both groups, however, higher titers were seen in previously symptomatic patients. There is a correlation between NAbs and both IgG/total anti-S1-SARS-CoV-2 antibodies (r = 0.47, P < 0.0001 and r = 0.49, P < 0.0001, respectively). IgG and total Ig could predict a neutralization titer of ≥ 1:160 at S/CO >4.44 and >65 with AUC 0.69 and 0.67, respectively. CONCLUSION: Asymptomatic SARS-CoV-2 infection can produce comparable antibodies response to previously symptomatic individuals, however higher neutralization activity was seen in the previously symptomatic. Anti-S1-SARS-CoV-2 IgG/total Ig antibodies showed a correlation with neutralization activity and can be used to estimate the presence of protective immunity.


Subject(s)
COVID-19 , SARS-CoV-2 , Antibodies, Neutralizing , Antibodies, Viral , COVID-19/diagnosis , Humans , Immunoassay , Immunoglobulin G , Luminescence
6.
J Mater Chem B ; 10(34): 6443-6452, 2022 08 31.
Article in English | MEDLINE | ID: covidwho-1890296

ABSTRACT

In this work, a designed porous DNA crystal with high intrinsic biocompatibility was used as the scaffold material to load fluorescent guest molecules to detect anti-cancer drugs. It is shown here that the synthesized crystals have the characteristics consistent with the designed large solvent channels, and can therefore accommodate guest molecules such as fluorescent proteins that cannot be accommodated by less porous crystals. Eu(TTA)3phen and Tb(acac)3phen lanthanide complexes were individually noncovalently loaded into the porous crystals, resulting in hybrid luminescent DNA crystals. Emodin, an anti-cancer, anti-tumor, anti-inflammatory drug, was found to quench lanthanide complexes in solution or in crystals. Notably, emodin is the active ingredient of Lianhua Qingwen Capsule, an anti-COVID-19 drug candidate. Therefore, the porous DNA crystals reported here have potential applications as a biocompatible and theranostic delivery biomaterial for functional macromolecules.


Subject(s)
Emodin , Lanthanoid Series Elements , DNA , Lanthanoid Series Elements/chemistry , Luminescence , Pharmaceutical Preparations
7.
Microbiol Spectr ; 10(3): e0103221, 2022 06 29.
Article in English | MEDLINE | ID: covidwho-1832366

ABSTRACT

This study includes 259 consecutive nasopharyngeal swabs which tested positive for a molecular SARS-CoV-2 test and 77 subjects who were followed longitudinally, with nasopharyngeal swabs performed weekly until clinical recovery and a negative result for the molecular test were reached. All swabs were also tested with a Lumipulse SARS-CoV-2 chemiluminescence enzyme immunoassay (CLEIA) antigen assay. The antigen test was positive in 169 (65.3%) out of the 259 subjects, while no antigen was detected in 90 subjects (34.7%). In the antigen-positive subjects, clinical status moved slightly toward a more frequent presence of symptoms. Longitudinal follow-up shows how the time of negativization has a faster kinetic in the antigenic test than in the molecular test. Antigenic test result values, considered as a time-dependent covariate and log-transformed, were highly associated with the time to negative swab, with good prediction ability. Receiver operating characteristic (ROC) curve analysis showed a very good discrimination ability of antigenic tests in classifying negative swabs. The optimal cutoff which jointly maximized sensitivity and specificity was 1.55, resulting in an overall accuracy of 0.75, a sensitivity of 0.73, and a specificity of 0.83. After dichotomizing the antigenic test according to the previously determined cutoff value of 1.55, the time-dependent covariate Cox model again suggests a highly significant association of antigenic test values with the time to negative swab molecular: a subject with an antigenic test value lower than 1.55 had almost a 13-fold higher probability to also result negative in the molecular test compared to a subject with an antigenic test value higher than 1.55. IMPORTANCE Our work explores the possibility of using a sensible and reliable antigenic test in a wider range of SARS-CoV-2 diagnostic and clinical applications. Furthermore, this tool seems particularly promising in follow-up with infected subjects, because while the molecular test frequently yields the persistence of low positivities, raising yet unanswered questions, this antigenic test shows more uniform and faster negativization during the evolution of the infection, somehow paralleling the dynamics of infectivity. Although more data will be required to definitely prove it, we believe these findings might be of great interest.


Subject(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnosis , Follow-Up Studies , Humans , Immunoenzyme Techniques , Luminescence , SARS-CoV-2/genetics
9.
J Clin Lab Anal ; 35(1): e23649, 2021 Jan.
Article in English | MEDLINE | ID: covidwho-1064371

ABSTRACT

OBJECTIVE: To explore the clinical value of serum IgM and IgG to SARS-CoV-2 in COVID-19. METHODS: 105 COVID-19 patients were enrolled as the disease group. 197 non-COVID-19 patients served as the control group. Magnetic chemiluminescent immunoassay (MCLIA) was used to detect the IgM and IgG. RESULTS: The peak of positive rates of SARS-CoV-2 IgM was about 1 week earlier than that of IgG. It reached to peak within 15-21 days and then began a slowly decline. The positive rates of IgG were increased with the disease course and reached the peak between 22 and 39 days. The differences in sensitivity of the three detection modes (IgM, IgG, and IgM + IgG) were statistically significant. The largest group of test cases (illness onset 15-21 days) showed that the positive rate of IgG was higher than IgM. Also, the sensitivity of IgM combined with IgG was higher than IgM or IgG. IgM and IgG were monitored dynamically for 16 patients with COVID-19, the results showed that serological transformation of IgM was carried out simultaneously with IgG in seven patients, which was earlier than IgG in four patients and later than IgG in five patients. CONCLUSION: The detection of SARS-CoV-2 IgM and IgG is very important to determine the course of COVID-19. Nucleic acid detection combined with serum antibody of SARS-CoV-2 may be the best laboratory indicator for the diagnosis of SARS-CoV-2 infection and the phrase and predication for prognosis of COVID-19.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , Immunoassay/methods , Immunoglobulin G/blood , Immunoglobulin M/blood , Adult , Antibodies, Viral/blood , Biomarkers/blood , Female , Humans , Luminescence , Male , Middle Aged , Retrospective Studies , SARS-CoV-2/genetics , SARS-CoV-2/immunology , Seroconversion
11.
Nature ; 591(7850): 482-487, 2021 03.
Article in English | MEDLINE | ID: covidwho-1049967

ABSTRACT

Naturally occurring protein switches have been repurposed for the development of biosensors and reporters for cellular and clinical applications1. However, the number of such switches is limited, and reengineering them is challenging. Here we show that a general class of protein-based biosensors can be created by inverting the flow of information through de novo designed protein switches in which the binding of a peptide key triggers biological outputs of interest2. The designed sensors are modular molecular devices with a closed dark state and an open luminescent state; analyte binding drives the switch from the closed to the open state. Because the sensor is based on the thermodynamic coupling of analyte binding to sensor activation, only one target binding domain is required, which simplifies sensor design and allows direct readout in solution. We create biosensors that can sensitively detect the anti-apoptosis protein BCL-2, the IgG1 Fc domain, the HER2 receptor, and Botulinum neurotoxin B, as well as biosensors for cardiac troponin I and an anti-hepatitis B virus antibody with the high sensitivity required to detect these molecules clinically. Given the need for diagnostic tools to track the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)3, we used the approach to design sensors for the SARS-CoV-2 spike protein and antibodies against the membrane and nucleocapsid proteins. The former, which incorporates a de novo designed spike receptor binding domain (RBD) binder4, has a limit of detection of 15 pM and a luminescence signal 50-fold higher than the background level. The modularity and sensitivity of the platform should enable the rapid construction of sensors for a wide range of analytes, and highlights the power of de novo protein design to create multi-state protein systems with new and useful functions.


Subject(s)
Antibodies, Viral/analysis , Biosensing Techniques/methods , Hepatitis B virus/immunology , SARS-CoV-2/chemistry , SARS-CoV-2/immunology , Spike Glycoprotein, Coronavirus/analysis , Troponin I/analysis , Antibodies, Viral/immunology , Biosensing Techniques/standards , Botulinum Toxins/analysis , Coronavirus Nucleocapsid Proteins/immunology , Immunoglobulin G/analysis , Immunoglobulin G/immunology , Limit of Detection , Luminescence , Phosphoproteins/immunology , Proto-Oncogene Proteins c-bcl-2/analysis , Receptor, ErbB-2/analysis , Sensitivity and Specificity , Viral Matrix Proteins/immunology
12.
Nano Lett ; 21(2): 1017-1024, 2021 01 27.
Article in English | MEDLINE | ID: covidwho-1028800

ABSTRACT

Bioaerosols, including infectious diseases such as COVID-19, are a continuous threat to global public safety. Despite their importance, the development of a practical, real-time means of monitoring bioaerosols has remained elusive. Here, we present a novel, simple, and highly efficient means of obtaining enriched bioaerosol samples. Aerosols are collected into a thin and stable liquid film by the unique interaction of a superhydrophilic surface and a continuous two-phase centrifugal flow. We demonstrate that this method can provide a concentration enhancement ratio of ∼2.4 × 106 with a collection efficiency of ∼99.9% and an aerosol-into-liquid transfer rate of ∼95.9% at 500 nm particle size (smaller than a single bacterium). This transfer is effective in both laboratory and external ambient environments. The system has a low limit of detection of <50 CFU/m3air using a straightforward bioluminescence-based technique and shows significant potential for air monitoring in occupational and public-health applications.


Subject(s)
Aerosols , Bacteria/isolation & purification , Environmental Monitoring/instrumentation , Environmental Monitoring/methods , Air Microbiology , Biomass , Limit of Detection , Luminescence , Nanoparticles , Particle Size , Public Health , Surface Properties , Temperature
13.
J Clin Lab Anal ; 35(1): e23681, 2021 Jan.
Article in English | MEDLINE | ID: covidwho-986204

ABSTRACT

BACKGROUND: Seldom performance evaluation and diagnosis comparison studies were reported for different chemiluminescent immunoassay (CLIA) kits approved under an emergency approval program for SARS-CoV-2 infection. METHODS: A total of 100 and 105 serum separately from non-infected populations and COVID-19 patients were detected with SARS-CoV-2 IgM and IgG kits. The characteristics including precision, functional sensitivity, linearity, and accuracy were evaluated for Axceed, iFlash, and Maglumi CLIA kits. RESULTS: Maglumi and iFlash had the best analytical sensitivity for IgM and IgG, respectively. Axceed kits had a linearity response in the detected concentration. The clinical sensitivity of Axceed, iFlash, and Maglumi was 68.0%, 64.9%, and 63.9% with a specificity of 99.0%, 96.0%, and 100% for IgM, 85.6%, 97.9%, and 94.8% with a specificity of 97.0% for IgG. ROC analysis indicated all kits had a diagnostic power greater than 0.9. Notably, either IgM or IgG kits obtained a poor agreement (Kappa value from 0.397 to 0.713) with others. Among 38 recovered patients, 94.7% had an effective immune response, and both seropositive IgM and IgG accounted for the biggest proportion (medium, 42 days onset), then followed by the single seropositive IgG (medium, 50 days onset) in Ab profile. CONCLUSION: The performance of CLIA kits satisfied the diagnosis of SARS-CoV-2 infection. Both positive of IgG and IgM contributes to improve the specificity, and a positive one will enhance the sensitivity.


Subject(s)
COVID-19 Testing/methods , COVID-19/etiology , Immunoassay/methods , Immunoglobulin G/blood , Immunoglobulin M/blood , Adult , Aged , Antibodies, Viral/blood , Automation, Laboratory , COVID-19/diagnosis , Female , Humans , Luminescence , Pregnancy , Pregnancy Complications, Infectious/etiology , Pregnancy Complications, Infectious/therapy , Reproducibility of Results , SARS-CoV-2/immunology , Time Factors
14.
Ann Clin Biochem ; 57(6): 429-434, 2020 11.
Article in English | MEDLINE | ID: covidwho-982712

ABSTRACT

OBJECTIVES: The purpose of the current study was to evaluate the analytical performance of seven kits for detecting IgM/IgG antibodies against coronavirus (SARS-CoV-2) by using four chemiluminescence immunoassay systems. METHODS: Fifty patients diagnosed with SARS-CoV-2 infection and 130 controls without coronavirus infection from the General Hospital of Chongqing were enrolled in the current retrospective study. Four chemiluminescence immunoassay systems, including seven IgM/IgG antibody detection kits for SARS-CoV-2 (A_IgM, A_IgG, B_IgM, B_IgG, C_IgM, C_IgG and D_Ab), were employed to detect antibody concentrations. The chi-square test, the receiver operating characteristic (ROC) curve and Youden's index were determined to verify the cut-off value of each detection system. RESULTS: The repeatability verification results of the A, B, C and D systems are all qualified. D_Ab performed best (92% sensitivity and 99.23% specificity), and B_IgM performed worse than the other systems. Except for the A_IgM and C_IgG systems, the optimal diagnostic thresholds and cut-off values of the other kits and their recommendations are inconsistent with each other. B_IgM had the worst AUC, and C_IgG had the best diagnostic accuracy. More importantly, the B_IgG system had the highest false-positive rate for testing patients with AIDS, tumours and pregnancies. The A_IgM system test showed the highest false-positive rates among elderly individuals over 90 years old. COVID-2019 IgM/IgG antibody test systems exhibit performance differences. CONCLUSIONS: The Innodx Biotech Total Antibody serum diagnosis kit is the most reliable detection system for anti-SARS-CoV-2 antibodies, which can be used together with nucleic acid tests as an alternative method for SARS-CoV-2 detecting.


Subject(s)
Antibodies, Viral/analysis , Antibody Specificity , Betacoronavirus/immunology , Immunoassay/methods , Luminescence , Antibodies, Viral/blood , Antibodies, Viral/immunology , Female , Humans , Limit of Detection , Male , Middle Aged , SARS-CoV-2
15.
J Med Virol ; 93(3): 1805-1809, 2021 03.
Article in English | MEDLINE | ID: covidwho-880291

ABSTRACT

Plasma specimens from coronavirus disease 2019 patients were double-tested for anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies by two different batches of MAGLUMI 2019-nCov immunoglobulin M/immunoglobulin G (IgM/IgG) assays to evaluate IgM/IgG levels, qualitative interpretation, antibody kinetics, and linearity of diluted specimen. Here we show that (i) high-level IgM specimens need to be diluted with negative human plasma but not kit diluents and (ii) measured anti-SARS-CoV-2 IgM/IgG concentrations are substantially higher with later marketed immunoassay batch leading to (iii) the change of qualitative interpretation (positive vs. negative) in 12.3% of specimens measured for IgM, (iv) the informative time-course pattern of antibody production only when data from different immunoassay batches are not combined.


Subject(s)
COVID-19/blood , COVID-19/immunology , Immunoglobulin G/blood , Immunoglobulin G/immunology , Immunoglobulin M/blood , Immunoglobulin M/immunology , SARS-CoV-2/immunology , Antibodies, Viral/immunology , COVID-19 Testing/methods , Humans , Immunoassay/methods , Luminescence , Luminescent Measurements/methods , Sensitivity and Specificity
16.
J Coll Physicians Surg Pak ; 30(8): 801-804, 2020 Aug.
Article in English | MEDLINE | ID: covidwho-745629

ABSTRACT

OBJECTIVE: To determine the percentage of seroconverted real time reverse transcriptase polymerase chain reaction (RT-PCR) positive coronavirus disease (COVID-19) cases at different days post-symptom onset; and also find the agreement of chemiluminescence assay used for total antibody detection using RT-PCR as a reference method. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Chughtai Institute of Pathology from April to May 2020. METHODOLOGY: Fifty pre-pandemic samples (healthy population) and 75 COVID-19 patients were included in the study. RT-PCR confirmed COVID-19 patients were divided into 3 equal groups (25 each), according to the days of symptom onset. The samples were analysed using electro-chemiluminescence as assay principle. Positive and negative agreement of COVID-19 antibodies was calculated using EP evaluator to find out the sensitivity of chemiluminescence assay for total antibody detection. The results were analysed using SPSS version 23.0. RESULTS: All the pre-pandemic samples tested were negative for antibodies with a negative agreement of 100%. Total agreement at day 7 post-symptom onset was 84%; whereas, it was 94% at day 14 and increased rapidly to 100% at day 21 post-symptom onset. At day 7 post-symptom onset, 68% of patients were seroconverted; and this percentage was 88% and 100% at day 14 and 21 post-symptom onset, respectively. CONCLUSION: Pre-pandemic samples were non-reactive for COVID-19 antibodies and seroconversion started within the first week post-virus exposure. There was 100% concordance between RT-PCR result and antibody positivity 21 days post-symptom onset. Key Words: COVID-19, SARS CoV-2, Seroconversion, Chemiluminescence.


Subject(s)
Antibodies, Viral/blood , Coronavirus Infections/diagnosis , Coronavirus/isolation & purification , Pneumonia, Viral/diagnosis , Adolescent , Adult , Betacoronavirus , COVID-19 , Coronavirus/genetics , Coronavirus/immunology , Coronavirus Infections/epidemiology , Cross-Sectional Studies , Female , Humans , Luminescence , Male , Middle Aged , Pakistan/epidemiology , Pandemics , Pneumonia, Viral/epidemiology , Real-Time Polymerase Chain Reaction , Risk Factors , SARS-CoV-2 , Seroconversion
17.
Biosens Bioelectron ; 166: 112436, 2020 Oct 15.
Article in English | MEDLINE | ID: covidwho-665846

ABSTRACT

Our recent experience of the COVID-19 pandemic has highlighted the importance of easy-to-use, quick, cheap, sensitive and selective detection of virus pathogens for the efficient monitoring and treatment of virus diseases. Early detection of viruses provides essential information about possible efficient and targeted treatments, prolongs the therapeutic window and hence reduces morbidity. Graphene is a lightweight, chemically stable and conductive material that can be successfully utilized for the detection of various virus strains. The sensitivity and selectivity of graphene can be enhanced by its functionalization or combination with other materials. Introducing suitable functional groups and/or counterparts in the hybrid structure enables tuning of the optical and electrical properties, which is particularly attractive for rapid and easy-to-use virus detection. In this review, we cover all the different types of graphene-based sensors available for virus detection, including, e.g., photoluminescence and colorimetric sensors, and surface plasmon resonance biosensors. Various strategies of electrochemical detection of viruses based on, e.g., DNA hybridization or antigen-antibody interactions, are also discussed. We summarize the current state-of-the-art applications of graphene-based systems for sensing a variety of viruses, e.g., SARS-CoV-2, influenza, dengue fever, hepatitis C virus, HIV, rotavirus and Zika virus. General principles, mechanisms of action, advantages and drawbacks are presented to provide useful information for the further development and construction of advanced virus biosensors. We highlight that the unique and tunable physicochemical properties of graphene-based nanomaterials make them ideal candidates for engineering and miniaturization of biosensors.


Subject(s)
Betacoronavirus/isolation & purification , Biosensing Techniques , Clinical Laboratory Techniques , Coronavirus Infections/diagnosis , Graphite , Pneumonia, Viral/diagnosis , Viruses/isolation & purification , Antigen-Antibody Reactions , Betacoronavirus/genetics , Betacoronavirus/pathogenicity , Biosensing Techniques/instrumentation , Biosensing Techniques/methods , Biosensing Techniques/trends , COVID-19 , COVID-19 Testing , Clinical Laboratory Techniques/instrumentation , Clinical Laboratory Techniques/methods , Clinical Laboratory Techniques/statistics & numerical data , Colorimetry , Coronavirus Infections/epidemiology , Coronavirus Infections/virology , DNA, Viral/analysis , DNA, Viral/genetics , Electrochemical Techniques , Equipment Design , Graphite/chemistry , Humans , Luminescence , Nanostructures/chemistry , Nucleic Acid Hybridization , Pandemics , Pneumonia, Viral/epidemiology , Pneumonia, Viral/virology , Quantum Dots/chemistry , SARS-CoV-2 , Spectrum Analysis, Raman , Surface Plasmon Resonance , Virology/methods , Viruses/genetics , Viruses/pathogenicity
19.
Clin Chem Lab Med ; 58(8): 1357-1364, 2020 Jul 28.
Article in English | MEDLINE | ID: covidwho-345901

ABSTRACT

Objectives Faced with the COVID-19 pandemic and its impact on the availability and quality of both therapeutic and diagnostic methods, the Belgian authorities have decided to launch a procedure for additional evaluation of the performance of serological tests offered for sale on the national territory. This has been proposed with a double aim: (1) an in-depth verification of the analytical and clinical performances presented by the manufacturer and (2) an economy of scale in terms of centralized validation for all the laboratories using the tests subject to evaluation. Methods A retrospective validation study was conducted including the serum of 125 patients in order to determine the analytical and clinical performances of the LIAISON®SARS-CoV-2 from DiaSorin® detecting anti-SARS-CoV-2 IgG and to compare its clinical performance with the enzyme-linked immunosorbent assay (ELISA) test from Euroimmun®, one of the first commercially available tests allowing the detection of anti-SARS-CoV-2 IgA and IgG. Results The performances of the LIAISON®SARS-CoV-2 satisfied all the acceptance criteria and provided "real world" analytical and clinical performances very close to the ones reported by the manufacturer in its insert kit. Comparison between the LIAISON®SARS-CoV-2 and the ELISA method did not reveal any difference between the two techniques in terms of sensitivities and specificities regarding the determination of the IgG. Conclusions This study reports the validation of the LIAISON®SARS-CoV-2 allowing to detect IgG antibodies specifically directed against SARS-CoV-2. The analytical and clinical performances are excellent, and the automation of the test offers important rates, ideal for absorbing an extension of testing.


Subject(s)
Antibodies, Viral/immunology , Betacoronavirus/immunology , Clinical Laboratory Techniques/methods , Coronavirus Infections/diagnosis , Immunoassay/methods , Pneumonia, Viral/diagnosis , COVID-19 , COVID-19 Testing , Coronavirus Infections/immunology , Humans , Limit of Detection , Luminescence , Luminescent Measurements/methods , Pandemics , Pneumonia, Viral/immunology , Reproducibility of Results , Retrospective Studies , SARS-CoV-2
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