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1.
Clin Pediatr (Phila) ; 61(1): 34-41, 2022 01.
Article in English | MEDLINE | ID: covidwho-1505941

ABSTRACT

In Italy, during the second epidemic wave of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), rapid antigenic (Ag) test at point-of-care (POCT) station were employed to quickly evaluate large numbers of swabs. We collected data of all children who underwent the Ag test in our hospital. All positive patients were recalled to perform reverse transcription polymerase chain reaction. A total of 2133 tests were collected over 1 month. Clinical data of 1941 children (median age = 3.7 years) were analyzed: 1343 (69.2%) patients complained of symptoms, 594 (30.6%) had a history of close contact with SARS-CoV-2-positive individuals. Among symptoms reported, acute rhinitis was the most frequent (67.9%), followed by cough (42.6%) and fever (31.5%). Among all tests, 95.8% resulted negative, 4.2% positive: 37/89 were confirmed. In confirmed cases, fever (56.2% vs 32.2%; P = .041) and gastrointestinal symptoms (18.8% vs 6.25%; P = .041) were significantly more frequent compared with negative children. The use of POCT for Ag test seems appropriate for SARS-CoV-2 screening in the pediatric population. In children, fever and gastrointestinal symptoms may constitute red flags of SARS-CoV-2.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , Point-of-Care Systems/standards , Adolescent , COVID-19/epidemiology , COVID-19 Testing/standards , COVID-19 Testing/statistics & numerical data , Child , Child, Preschool , Female , Humans , Italy/epidemiology , Male , Pediatrics/methods , Point-of-Care Systems/statistics & numerical data , Young Adult
4.
PLoS One ; 16(9): e0257464, 2021.
Article in English | MEDLINE | ID: covidwho-1416902

ABSTRACT

Despite the development of effective vaccines against SARS-CoV-2, epidemiological control of the virus is still challenging due to slow vaccine rollouts, incomplete vaccine protection to current and emerging variants, and unwillingness to get vaccinated. Therefore, frequent testing of individuals to identify early SARS-CoV-2 infections, contact-tracing and isolation strategies remain crucial to mitigate viral spread. Here, we describe WHotLAMP, a rapid molecular test to detect SARS-CoV-2 in saliva. WHotLAMP is simple to use, highly sensitive (~4 viral particles per microliter of saliva) and specific, as well as inexpensive, making it ideal for frequent screening. Moreover, WHotLAMP does not require toxic chemicals or specialized equipment and thus can be performed in point-of-care settings, and may also be adapted for resource-limited environments or home use. While applied here to SARS-CoV-2, WHotLAMP can be modified to detect other pathogens, making it adaptable for other diagnostic assays, including for use in future outbreaks.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19/diagnosis , RNA, Viral/genetics , SARS-CoV-2/genetics , Saliva/virology , COVID-19/epidemiology , COVID-19/virology , COVID-19 Nucleic Acid Testing/instrumentation , Epidemics/prevention & control , Humans , Point-of-Care Systems/statistics & numerical data , RNA, Viral/isolation & purification , Reproducibility of Results , SARS-CoV-2/physiology , Sensitivity and Specificity
5.
Biosens Bioelectron ; 178: 113049, 2021 Apr 15.
Article in English | MEDLINE | ID: covidwho-1056383

ABSTRACT

Prompt diagnosis, patient isolation, and contact tracing are key measures to contain the coronavirus disease 2019 (COVID-19). Molecular tests are the current gold standard for COVID-19 detection, but are carried out at central laboratories, delaying treatment and control decisions. Here we describe a portable assay system for rapid, onsite COVID-19 diagnosis. Termed CODA (CRISPR Optical Detection of Anisotropy), the method combined isothermal nucleic acid amplification, activation of CRISPR/Cas12a, and signal generation in a single assay, eliminating extra manual steps. Importantly, signal detection was based on the ratiometric measurement of fluorescent anisotropy, which allowed CODA to achieve a high signal-to-noise ratio. For point-of-care operation, we built a compact, standalone CODA device integrating optoelectronics, an embedded heater, and a microcontroller for data processing. The developed system completed SARS-CoV-2 RNA detection within 20 min of sample loading; the limit of detection reached 3 copy/µL. When applied to clinical samples (10 confirmed COVID-19 patients; 10 controls), the rapid CODA test accurately classified COVID-19 status, in concordance with gold-standard clinical diagnostics.


Subject(s)
Biosensing Techniques/methods , COVID-19 Nucleic Acid Testing/methods , COVID-19/diagnosis , Fluorescence Polarization/methods , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Biosensing Techniques/instrumentation , Biosensing Techniques/statistics & numerical data , COVID-19/virology , COVID-19 Nucleic Acid Testing/instrumentation , COVID-19 Nucleic Acid Testing/statistics & numerical data , CRISPR-Cas Systems , Equipment Design , Fluorescence Polarization/instrumentation , Fluorescence Polarization/statistics & numerical data , Humans , Molecular Diagnostic Techniques/instrumentation , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/statistics & numerical data , Nucleic Acid Amplification Techniques/instrumentation , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/statistics & numerical data , Pandemics , Point-of-Care Systems/statistics & numerical data , Signal Processing, Computer-Assisted , Signal-To-Noise Ratio
6.
Biosens Bioelectron ; 178: 113041, 2021 Apr 15.
Article in English | MEDLINE | ID: covidwho-1051492

ABSTRACT

The outbreak of COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been challenging human health worldwide. Loop-mediated isothermal amplification (LAMP) has been promptly applied to the detection of SARS-CoV-2 owing to its high amplification efficacy and less requirement of the thermal cycler. However, the vast majority of these LAMP-based assays depend on the non-specific detection of LAMP products, which can not discern the undesirable amplificons, likely to yield unreliable results. Herein, a sequence-specific LAMP assay was reported to detect SARS-CoV-2 using proofreading enzyme-mediated probe cleavage (named Proofman), which could realize real-time and visual detection without uncapping. This assay, introducing a proofreading enzyme and the fluorogenic probe to reverse-transcription LAMP (RT-Proofman-LAMP), can specifically detect the SARS-CoV-2 RNA with a detection limit of 100 copies. In addition to the real-time analysis, the assay is capable of endpoint visualization under a transilluminator within 50 min, providing a convenient reporting manner under the setting of point-of-care testing (POCT). In combination with different fluorophores, the one-pot multiplex assay was successfully achieved to detect multiple targets of SARS-CoV-2 and inner control simultaneously. In summary, the development of RT-Proofman-LAMP offers a versatile and highly-specific method for fast field screening and laboratory testing of SARS-CoV-2, making it a promising platform in COVID-19 diagnosis.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19/diagnosis , COVID-19/virology , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Biosensing Techniques/methods , Biosensing Techniques/statistics & numerical data , COVID-19 Nucleic Acid Testing/statistics & numerical data , Humans , Limit of Detection , Molecular Diagnostic Techniques/statistics & numerical data , Multiplex Polymerase Chain Reaction/methods , Multiplex Polymerase Chain Reaction/statistics & numerical data , Nucleic Acid Amplification Techniques/statistics & numerical data , Pandemics , Point-of-Care Systems/statistics & numerical data , RNA, Viral/analysis , RNA, Viral/genetics , Sensitivity and Specificity
7.
J Transl Med ; 18(1): 488, 2020 12 21.
Article in English | MEDLINE | ID: covidwho-992499

ABSTRACT

BACKGROUND: The easy access to a quick diagnosis of coronavirus disease 2019 (COVID-19) is a key point to improve the management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and to contain its spread. Up to now, laboratory real-time PCR is the standard of care, but requires a fully equipped laboratory and significant infrastructure. Consequently, new diagnostic tools are required. METHODS: In the present work, the diagnostic accuracy of the point-of-care rapid test "bKIT Virus Finder COVID-19" (Hyris Ltd) is evaluated by a retrospective and a prospective analysis on SARS CoV-2 samples previously assessed with an FDA "authorized for the emergency use-EUA" reference method. Descriptive statistics were used for the present study. RESULTS: Results obtained with the Hyris Kit are the same as that of standard laboratory-based real time PCR methods for all the analyzed samples. In addition, the Hyris Kit provides the test results in less than 2 h, a significantly shorter time compared to the reference methods, without the need of a fully equipped laboratory. CONCLUSIONS: To conclude, the Hyris kit represents a promising tool to improve the health surveillance and to increase the capacity of SARS-CoV-2 testing.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19/diagnosis , Point-of-Care Systems , COVID-19/epidemiology , COVID-19/virology , COVID-19 Nucleic Acid Testing/standards , COVID-19 Nucleic Acid Testing/statistics & numerical data , Early Diagnosis , Humans , Italy/epidemiology , Limit of Detection , Pandemics , Point-of-Care Systems/standards , Point-of-Care Systems/statistics & numerical data , Prospective Studies , Reference Standards , Retrospective Studies , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Sensitivity and Specificity
8.
Crit Care ; 24(1): 676, 2020 12 07.
Article in English | MEDLINE | ID: covidwho-962957

ABSTRACT

BACKGROUND: There is emerging evidence for enhanced blood coagulation in coronavirus 2019 (COVID-19) patients, with thromboembolic complications contributing to morbidity and mortality. The mechanisms underlying this prothrombotic state remain enigmatic. Further data to guide anticoagulation strategies are urgently required. METHODS: We used viscoelastic rotational thromboelastometry (ROTEM) in a single-center cohort of 40 critically ill COVID-19 patients. RESULTS: Clear signs of a hypercoagulable state due to severe hypofibrinolysis were found. Maximum lysis, especially following stimulation of the extrinsic coagulation system, was inversely associated with an enhanced risk of thromboembolic complications. Combining values for maximum lysis with D-dimer concentrations revealed high sensitivity and specificity of thromboembolic risk prediction. CONCLUSIONS: The study identifies a reduction in fibrinolysis as an important mechanism in COVID-19-associated coagulopathy. The combination of ROTEM and D-dimer concentrations may prove valuable in identifying patients requiring higher intensity anticoagulation.


Subject(s)
COVID-19/complications , Fibrinolysis/physiology , Thrombelastography/methods , Thromboembolism/diagnosis , Blood Coagulation/physiology , Blood Coagulation Tests/methods , Blood Coagulation Tests/standards , COVID-19/diagnostic imaging , COVID-19/physiopathology , Cohort Studies , Critical Illness/epidemiology , Critical Illness/therapy , Female , Humans , Male , Middle Aged , Point-of-Care Systems/standards , Point-of-Care Systems/statistics & numerical data , Thromboembolism/diagnostic imaging , Viscoelastic Substances/analysis , Viscoelastic Substances/therapeutic use
9.
Ann Biol Clin (Paris) ; 78(4): 363-382, 2020 08 01.
Article in French | MEDLINE | ID: covidwho-709296

ABSTRACT

The point-of-care tests (POCT) are subject to accreditation. A national inventory survey provides a synthesis of knowledge. The survey distributed 31 questions in 2019. 147 responses were received (75% biologists, 49% CHU, 42% CHG). Only 20.41% are accredited ISO22870, the majority for <50% of the medical departments; 70% say they are going there at the end of 2019 or in 2020. The maps are unknown for 32% (EBMD) and 82% (TROD). Visibility is poor with: medical establishment committee (40%), IT department (31%). Connection is necessary for 87-95% depending on the criterion (QC, authorizations, etc.) and 66% of answers highlight that less than 50% of connexion is effective. The major advantage is the delay of the result (62.5%), then the relationship with the health teams (33.3%). The disadvantages: difficulty of the quality approach (45%), cost of tests (34.3%). Human resource requirements are identified for technicians (82%) and biologists (76%). The multiplicity of sites, devices and operators means that it is difficult to set up and maintain. Biology outside the laboratories, under biological responsibility, must meet a rigorous imperative quality approach.


Subject(s)
Clinical Laboratory Techniques , Global Health , Laboratories/statistics & numerical data , Laboratories/standards , Point-of-Care Testing , Accreditation , COVID-19 , COVID-19 Testing , Clinical Laboratory Services/standards , Clinical Laboratory Services/statistics & numerical data , Clinical Laboratory Techniques/standards , Clinical Laboratory Techniques/statistics & numerical data , Coronavirus Infections/diagnosis , Coronavirus Infections/epidemiology , France/epidemiology , Global Health/standards , Global Health/statistics & numerical data , History, 21st Century , Humans , Internationality , Laboratory Proficiency Testing/standards , Pandemics , Pneumonia, Viral/diagnosis , Pneumonia, Viral/epidemiology , Point-of-Care Systems/standards , Point-of-Care Systems/statistics & numerical data , Point-of-Care Testing/organization & administration , Point-of-Care Testing/standards , Point-of-Care Testing/statistics & numerical data , Quality Assurance, Health Care/organization & administration , Surveys and Questionnaires
10.
J Appl Lab Med ; 5(6): 1307-1312, 2020 11 01.
Article in English | MEDLINE | ID: covidwho-696741

ABSTRACT

BACKGROUND: Numerous nucleic acid amplification assays utilizing different target genes of the SARS-CoV-2 genome have received emergency use authorization (EUA) by the United States Food and Drug Administration (FDA). Limited data are available comparing the test performance characteristics of these assays. METHODS: A diagnostic comparison study was performed to evaluate the performance of the Cepheid Xpert Xpress SARS-CoV-2 assay compared to the Hologic Panther Fusion SARS-CoV-2 assay using clinical nasopharyngeal specimens. Agreement between the two assays was assessed by overall, positive, and negative percent agreement and Cohen's kappa coefficient. RESULTS: A total of 104 (54 positive and 50 negative) clinical nasopharyngeal samples were tested by both assays. Using the Panther Fusion as a reference standard, the Xpert demonstrated an overall agreement of 99.0% [95% confidence interval (CI): 94.8-100], positive percent agreement of 98.1% (95% CI: 90.1-100), and a negative percent agreement of 100% (95% CI: 94.2-100). The kappa coefficient was 0.98 (95% CI: 0.94-1.0). One sample positive by the Panther Fusion with a cycle threshold (Ct) of 38.6 was found to be reproducibly negative by the Xpert assay. CONCLUSIONS: The Cepheid Xpert Xpress SARS-CoV-2 assay provides test performance comparable to the Hologic Panther Fusion SARS-CoV-2 assay while offering laboratories rapid, on-demand testing capacity.


Subject(s)
Betacoronavirus/isolation & purification , Clinical Laboratory Techniques/instrumentation , Coronavirus Infections/diagnosis , Molecular Diagnostic Techniques/instrumentation , Pneumonia, Viral/diagnosis , RNA, Viral/isolation & purification , Automation, Laboratory/instrumentation , Betacoronavirus/genetics , COVID-19 , COVID-19 Testing , Clinical Laboratory Techniques/statistics & numerical data , Coronavirus Infections/epidemiology , Coronavirus Infections/virology , Humans , Molecular Diagnostic Techniques/statistics & numerical data , Multiplex Polymerase Chain Reaction/instrumentation , Pandemics , Pneumonia, Viral/epidemiology , Pneumonia, Viral/virology , Point-of-Care Systems/statistics & numerical data , Reagent Kits, Diagnostic/statistics & numerical data , Real-Time Polymerase Chain Reaction/instrumentation , Reproducibility of Results , SARS-CoV-2 , Time Factors , United States/epidemiology
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