scholarly journals Comparative performance of SARS CoV-2 lateral flow antigen tests demonstrates their utility for high sensitivity detection of infectious virus in clinical specimens

2021 ◽  
Author(s):  
Suzanne Pickering ◽  
Rahul Batra ◽  
Luke B. Snell ◽  
Blair Merrick ◽  
Gaia Nebbia ◽  
...  
Keyword(s):  
Infectious Virus ◽  
Rapid Test ◽  
Lateral Flow ◽  
Clinical Samples ◽  
Successful Implementation ◽  
Virus Infectivity ◽  
Repeated Testing ◽  
Comparative Performance ◽  
Virus Culture ◽  
Antigen Tests

AbstractBackgroundRapid antigen lateral flow devices (LFDs) are set to become a cornerstone of SARS-CoV-2 mass community testing. However, their reduced sensitivity compared to PCR has raised questions of how well they identify infectious cases. Understanding their capabilities and limitations is therefore essential for successful implementation. To address this, we evaluated six commercial LFDs on the same collection of clinical samples and assessed their correlation with infectious virus culture and cycle threshold (Ct) values.MethodsA head-to-head comparison of specificities and sensitivities was performed on six commercial rapid antigen tests using combined nasal/oropharyngeal swabs, and their limits of detection determined using viral plaque forming units (PFU). Three of the LFDs were selected for a further study, correlating antigen test result with RT-PCR Ct values and positive viral culture in Vero-E6 cells. This included sequential swabs and matched serum samples obtained from four infected individuals with varying disease severities. Detection of antibodies was performed using an IgG/IgM Rapid Test Cassette, and neutralising antibodies by infectious virus assay. Finally, the sensitivities of selected rapid antigen LFTs were assessed in swabs with confirmed B.1.1.7 variant, currently the dominant genotype in the UK.FindingsMost of the rapid antigen LFDs showed a high specificity (>98%), and accurately detected 50 PFU/test (equivalent N1 Ct of 23.7 or RNA copy number of 3×106/ml). Sensitivities of the LFDs performed on clinical samples ranged from 65 to 89%. These sensitivities increased in most tests to over 90% for samples with Cts lower than 25. Positive virus culture was achieved for 57 out of 141 samples, with 80% of the positive cultures from swabs with Cts lower than 23. Importantly, sensitivity of the LFDs increased to over 95% when compared with the detection of infectious virus alone, irrespective of Ct. Longitudinal studies of PCR-positive samples showed that most of the tests identified all infectious samples as positive, but differences in test sensitivities can lead to missed cases in the absence of repeated testing. Finally, test performance was not impacted when re-assessed against swabs positive for the dominant UK variant B.1.1.7.InterpretationIn this comprehensive comparison of antigen LFD and virus infectivity, we demonstrate a clear relationship between Ct values, quantitative culture of infectious virus and antigen LFD positivity in clinical samples. Our data support regular testing of target groups using LFDs to supplement the current PCR testing capacity, to rapidly identify infected individuals in situations where they would otherwise go undetected.FundingKing’s Together Rapid COVID-19, Medical Research Council, Wellcome Trust, Huo Family Foundation.

A prospective nationwide observational study of agreement of antigen tests on oral pharyngeal swabs or less invasive testing with RT-qPCR, for detecting SARS-CoV-2 in adults: Protocol description (Preprint)

2021 ◽  
Author(s):  
Uffe Vest Schneider ◽  
Jenny Dahl Knudsen ◽  
Anders Koch ◽  
Nikolai Søren Kirkby ◽  
Jan Gorm Lisby
Keyword(s):  
Confidence Intervals ◽  
Predictive Value ◽  
Large Scale ◽  
Reference Method ◽  
Lateral Flow ◽  
Clinical Samples ◽  
Sampling Location ◽  
Analytical Sensitivity ◽  
Clinical Sensitivity ◽  
Antigen Tests

BACKGROUND The SARS-CoV-2 pandemic has resulted in an unprecedented level of world-wide testing for epidemiologic and diagnostic purposes, and due to the extreme need for tests, the gold standard reverse transcription polymerase chain reaction (RT-qPCR) testing capacity has been unable to meet the overall global testing demand. Consequently, although current literature has shown the sensitivity of rapid antigen tests (RATs) to be inferior to RT-qPCR, RATs have been implemented on a large scale without solid data on performance. OBJECTIVE This study will compare analytical and clinical sensitivities and specificities of 50 lateral flow or laboratory based RATs and three Strand Invasion Based Amplification (SIBA)-rt-PCR tests from 30 manufacturers to RT-qPCR on samples obtained from the deep oropharynx. In addition, the study will compare sensitivities and specificities of the included RATs as well as RT-qPCR on clinical samples obtained from the deep oropharynx, anterior nasal cavity, saliva, deep nasopharynx and expired air to RT-qPCR from deep oropharyngeal samples. METHODS In the prospective part of the study, 200 individuals found SARS-CoV-2 positive and 200 individuals found SARS-CoV-2 negative by routine RT-qPCR testing will be re-tested with each RAT applying RT-qPCR as the reference method. In the retrospective part of the study, 304 deep oropharyngeal cavity swabs divided into four groups based on RT-qPCR Cq levels will be tested by each RAT. RESULTS The results will be reported in several manuscripts with different aims. The first manuscript will report retrospective (analytical sensitivity, overall and stratified into different Cq range groups) and prospective (clinical sensitivity) data for RATs with RT-qPCR results as the reference method. The second manuscript will report results for RAT based on anatomical sampling location. The third manuscript will compare different anatomical sampling locations by RT-qPCR testing. The fourth manuscript will focus on RATs that rely on central laboratory testing. Test from four different manufactures will be compared for analytical performance data on retrospective deep oropharyngeal swab samples. The fifth manuscript will report the results of four RATs applied both as professional use and as self-test. The last manuscript will report the results from two breath tests participating in the study. Comparison of sensitivity and specificity between RATs will be done using McNemar for paired samples and chi-squared test for unpaired samples. Comparison of PPV and NPV between RATs will be done by bootstrap test. 95 % confidence intervals for sensitivity, specificity, positive predictive value and negative predictive value are calculated as bootstrap confidence intervals CONCLUSIONS The study will compare the sensitivities of a large number of RATs for SARS-CoV-2 compared to RT-qPCR and will address whether lateral flow based RATs test differ significantly from laboratory based RATS. The anatomical test location for both RAT and RT-qPCR will be compared. CLINICALTRIAL ClinicalTrials.gov NCT04913116

scholarly journals Comparative Sensitivity of Rapid Antigen Tests for the Delta Variant (B.1.617.2) of SARS-CoV-2

Viruses ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 2183
Author(s):  
Yuko Sakai-Tagawa ◽  
Seiya Yamayoshi ◽  
Peter J. Halfmann ◽  
Yoshihiro Kawaoka
Keyword(s):  
Rapid Detection ◽  
Influenza A ◽  
Infectious Virus ◽  
Rapid Test ◽  
Rapid Diagnosis ◽  
Antigen Test ◽  
Detection Level ◽  
Test Kit ◽  
Low Levels ◽  
Antigen Tests

Rapid antigen tests (RATs) for COVID-19 based on lateral flow immunoassays are useful for rapid diagnosis in a variety of settings. Although many kinds of RATs are available, their respective sensitivity has not been compared. Here, we examined the sensitivity of 27 RATs available in Japan for the detection of the SARS-CoV-2 delta variant. All of the RATs tested detected the delta variant albeit with different sensitivities. Nine RATs (ESPLINE SARS-CoV-2, ALSONIC COVID-19 Ag, COVID-19 and Influenza A+B Antigen Combo Rapid Test, ImmunoArrow SARS-CoV-2, Fuji Dri-chem immuno AG cartridge COVID-19 Ag, 2019-nCoV Ag rapid detection kit, Saliva SARS-CoV-2(2019-nCoV) Antigen Test Kit, and Rabliss SARS-CoV-2 antigen detection kit COVID19 AG) showed superior sensitivity to the isolated delta variant. Although actual clinical specimens were not examined, the detection level of most of the RATs was 7500 pfu, indicating that individuals whose test samples contained less virus than that would be considered negative. Therefore, it is important to bear in mind that RATs may miss individuals shedding low levels of infectious virus.

scholarly journals Comparison of Rapid Antigen Tests for COVID-19

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1420
Author(s):  
Seiya Yamayoshi ◽  
Yuko Sakai-Tagawa ◽  
Michiko Koga ◽  
Osamu Akasaka ◽  
Ichiro Nakachi ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Quantitative Pcr ◽  
Reverse Transcription ◽  
Infectious Virus ◽  
Virus Isolation ◽  
Rapid Diagnosis ◽  
Lateral Flow ◽  
Viral Antigens ◽  
Reverse Transcription Quantitative Pcr ◽  
Antigen Tests

Reverse transcription-quantitative PCR (RT-qPCR)-based tests are widely used to diagnose coronavirus disease 2019 (COVID-19). As a result that these tests cannot be done in local clinics where RT-qPCR testing capability is lacking, rapid antigen tests (RATs) for COVID-19 based on lateral flow immunoassays are used for rapid diagnosis. However, their sensitivity compared with each other and with RT-qPCR and infectious virus isolation has not been examined. Here, we compared the sensitivity among four RATs by using severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) isolates and several types of COVID-19 patient specimens and compared their sensitivity with that of RT-qPCR and infectious virus isolation. Although the RATs read the samples containing large amounts of virus as positive, even the most sensitive RAT read the samples containing small amounts of virus as negative. Moreover, all RATs tested failed to detect viral antigens in several specimens from which the virus was isolated. The current RATs will likely miss some COVID-19 patients who are shedding infectious SARS-CoV-2.

scholarly journals Diagnostic accuracy of Panbio™ rapid antigen tests on oropharyngeal swabs for detection of SARS-CoV-2

2021 ◽  
Author(s):  
Marie Thérèse Ngo Nsoga ◽  
Ilona Kronig ◽  
Francisco Javier Perez Rodriguez ◽  
Pascale Sattonnet-Roche ◽  
Diogo Da Silva ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Infectious Virus ◽  
Reference Sample ◽  
Rapid Test ◽  
Rapid Diagnostic Tests ◽  
Sample Type ◽  
Threshold Values ◽  
Reverse Transcription Quantitative Pcr ◽  
Antigen Tests ◽  
A Prospective Study

AbstractBackgroundAntigen-detecting rapid diagnostic tests (Ag-RDTs) for the detection of SARS-CoV-2 offer new opportunities for testing in the context of the COVID-19 pandemic. Nasopharyngeal swabs (NPS) are the reference sample type, but oropharyngeal swabs (OPS) may be a more acceptable sample type in some patients.MethodsWe conducted a prospective study in a single screening center to assess the diagnostic performance of the Panbio™ COVID-19 Ag Rapid Test (Abbott) on OPS compared with reverse-transcription quantitative PCR (RT-qPCR) using NPS.Results402 outpatients were enrolled in a COVID-19 screening center, of whom 168 (41.8%) had a positive RT-qPCR test. The oropharyngeal Ag-RDT sensitivity compared to nasopharyngeal RT-qPCR was 81% (95%CI: 74.2-86.6). Two false positives were noted out of the 234 RT-qPCR negative individuals, which resulted in a specificity of 99.1% (95%CI: 96.9-99.9) for the Ag-RDT.For cycle threshold values ≤ 26.7 (≥ 1E6 SARS-CoV-2 genomes copies/mL, a presumed cut-off for infectious virus), 96.3% sensitivity (95%CI: 90.7-99.0%) was obtained with the Ag-RDT using OPS.InterpretationBased on our findings, the diagnostic performance of the Panbio™ Covid-19 RDT with OPS samples meet the criteria required by the WHO for Ag-RDTs (sensitivity≥80% and specificity ≥97%).

scholarly journals The sensitivity of SARS-CoV-2 antigen tests in the view of large-scale testing

2020 ◽  
Author(s):  
Pavel Drevinek ◽  
Jakub Hurych ◽  
Zdenek Kepka ◽  
Ales Briksi ◽  
Michal Kulich ◽  
...  
Keyword(s):  
Large Scale ◽  
Point Of Care ◽  
Rapid Test ◽  
Antigen Test ◽  
Repeated Testing ◽  
Diagnostic Pcr ◽  
Large Scale Testing ◽  
Oropharyngeal Swab ◽  
Antigen Tests ◽  
Low Sensitivity

AbstractObjectivesAntigen tests have recently emerged as an interesting alternative to SARS-CoV-2 diagnostic PCR, thought to be valuable especially for the screening of bigger communities. To check appropriateness of the antigen based testing, we determined sensitivity of two point-of-care antigen tests when applied to a cohort of COVID-19 symptomatic, COVID-19 asymptomatic and healthy persons.MethodsWe examined nasopharyngeal swabs with antigen test 1 (Panbio Covid-19 Ag Rapid Test, Abbott) and antigen test 2 (Standard F Covid-19 Ag FIA, SD Biosensor). An additional nasopharyngeal and oropharyngeal swab of the same individual was checked with PCR (Allplex SARS-nCoV-2, Seegene). Within a 4-day period in October 2020, we collected specimens from 591 subjects. Of them, 290 had COVID-19 associated symptoms.ResultsWhile PCR positivity was detected in 223 cases, antigen test 1 and antigen test 2 were found positive in 148 (sensitivity 0.664, 95% CI 0.599 - 0.722) and 141 (sensitivity 0.623, 95% CI 0.558 - 0.684) patients, respectively. When only symptomatic patients were analysed, sensitivity increased to 0.738 (95% CI 0.667 - 0.799) for the antigen test 1 and to 0.685 (95% CI 0.611 - 0.750) for the antigen test 2. The substantial drop in sensitivity to 12.9% (95% CI 0.067 - 0.234) was observed for samples with the PCR threshold cycle above > 30.ConclusionsLow sensitivity of antigen tests leads to the considerable risk of false negativity. It is advisable to implement repeated testing with high enough frequency if the antigen test is used as a frontline screening tool.

scholarly journals Comparison of seven commercial SARS-CoV-2 rapid Point-of-Care Antigen tests

2020 ◽  
Author(s):  
Victor M. Corman ◽  
Verena Claudia Haage ◽  
Tobias Bleicker ◽  
Marie Luisa Schmidt ◽  
Barbara Mühlemann ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Rapid Test ◽  
Infectious Period ◽  
Clinical Samples ◽  
Virus Concentration ◽  
Viral Loads ◽  
And Performance ◽  
Antigen Tests ◽  
Point Of Care Tests

AbstractBackgroundAntigen point of care tests (AgPOCT) can accelerate SARS-CoV-2 testing. As first AgPOCT are becoming available, there is a growing interest in their utility and performance.MethodsHere we compare AgPOCT products by seven suppliers: the Abbott Panbio™ COVID-19 Ag Rapid Test; the RapiGEN BIOCREDIT COVID-19 Ag; the Healgen® Coronavirus Ag Rapid Test Cassette (Swab); the Coris Bioconcept Covid.19 Ag Respi-Strip; the R-Biopharm RIDA®QUICK SARS-CoV-2 Antigen; the NAL von minden NADAL COVID19-Ag Test; and the Roche/SD Biosensor SARS-CoV Rapid Antigen Test. Tests were evaluated on recombinant nucleoprotein, cultured endemic and emerging coronaviruses, stored clinical samples with known SARS-CoV-2 viral loads (n=138), stored samples from patients with respiratory agents other than SARS-CoV-2 (n=100), as well as self-sampled swabs from healthy volunteers (n=35).FindingsLimits of detection in six of seven tested products ranged between 2.08 × 106 and 2.88 × 107 copies per swab, the outlier at 1.58 × 1010 copies per swab. Specificities ranged between 98.53% and 100% in five products, with two outliers at 94.85% and 88.24%. False positive results were not associated with any specific respiratory agent. As some of the tested AgPOCT were early production lots, the observed issues with specificity are unlikely to persist.InterpretationThe sensitivity range of most AgPOCT overlaps with viral load figures typically observed during the first week of symptoms, which marks the infectious period in the majority patients. AgPOCTs with a limit of detection that approximates the virus concentration above which patients are infectious may enable shortcuts in decision-making in various areas of healthcare and public health.

scholarly journals Lateral flow antigen tests can sensitively detect live cultured virus of the SARS-CoV-2 B.1.1.7 lineage

2021 ◽  
Author(s):  
Konstantina Kontogianni ◽  
Ana I. Cubas-Atienzar ◽  
Dominic Wooding ◽  
Kate Buist ◽  
Caitlin R. Thompson ◽  
...  
Keyword(s):  
Lateral Flow ◽  
Antigen Tests

Evaluation of rapid test kits for quantification of HT-2 and T-2 toxins in naturally contaminated oats

2013 ◽  
Vol 6 (1) ◽  
pp. 31-41 ◽  
Author(s):  
H.U. Aamot ◽  
I.S. Hofgaard ◽  
G. Brodal ◽  
O. Elen ◽  
B. Holen ◽  
...  
Keyword(s):  
Concentration Ratio ◽  
Rapid Test ◽  
Reference Method ◽  
Lateral Flow ◽  
Cross Reaction ◽  
Cross Reactions ◽  
Lateral Flow Device ◽  
Simultaneous Testing ◽  
Flow Device ◽  
Mycotoxin Analysis

The aim of this study was to evaluate the performance and usefulness of three rapid test kits for analysis of HT-2 and T-2 toxins (HT-2 and T-2), two of the most potent trichothecenes commonly found in European oats. Concentrations of these two toxins combined (HT-2+T-2) were analysed in naturally contaminated oat samples (n=68) using the following test kits: Ridascreen® FAST T-2 Toxin (‘Fast ELISA’), DRAFT Ridascreen® HT-2/T-2 (‘Standard ELISA’, not commercially available), and the lateral flow device ROSA® HT-2-T-2 (‘Rosa LFD’). Mycotoxin analysis by LC-MS/MS was used as a reference method. Rosa LFD offered the best reliability, achieving detection that was stable across toxin levels, whereas detection by both ELISA kits differed significantly among toxin levels (P<0.01). The kits were also evaluated regarding agreement with the reference method (measured as Cohen's kappa) at a HT-2+T-2 concentration of 1000 μg/kg in naturally contaminated oats. Agreement was greatest for Rosa LFD (89.2%), intermediate for Standard ELISA (66.8%), and lowest for Fast ELISA (62.2%). Rosa LFD showed cross-reaction of 100% with both T-2 and HT-2. For the ELISA kits, cross-reactions were 100% with T-2 but below 100% with HT-2. Therefore, to estimate the sum of HT-2 and T-2 in an oat sample, it was necessary to re-calculate the data from both ELISA kits according to the known cross-reaction of each kit with HT-2 and the concentration ratio of HT-2 to T-2 in Norwegian oats. Rosa LFD had the highest correlation with LC-MS/MS (R2=0.94), and the corresponding R2 values for Fast and Standard ELISA were 0.61 and 0.83, respectively. Rosa LFD was well suited for on-site detection. Standard ELISA allows simultaneous testing of several samples that are useful for centralised laboratories.

scholarly journals Quantitative measurement of infectious virus in SARS-CoV-2 Alpha, Delta and Epsilon variants reveals higher infectivity (viral titer:RNA ratio) in clinical samples containing the Delta and Epsilon variants.

2021 ◽  
Author(s):  
Hannah W Despres ◽  
Margaret G Mills ◽  
David J Shirley ◽  
Madaline M Schmidt ◽  
Meei-Li Huang ◽  
...  
Keyword(s):  
Quantitative Measurement ◽  
Infectious Virus ◽  
Viral Rna ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Genome Copy ◽  
Live Virus ◽  
High Degree ◽  
The Relationship ◽  
Rna Levels

ABSTRACT Background Novel SARS-CoV-2 Variants of Concern (VoC) pose a challenge to controlling the COVID-19 pandemic. Previous studies indicate that clinical samples collected from individuals infected with the Delta variant may contain higher levels of RNA than previous variants, but the relationship between viral RNA and infectious virus for individual variants is unknown. Methods We measured infectious viral titer (using a micro-focus forming assay) as well as total and subgenomic viral RNA levels (using RT-PCR) in a set of 165 clinical samples containing SARS-CoV-2 Alpha, Delta and Epsilon variants that were processed within two days of collection from the patient. Results We observed a high degree of variation in the relationship between viral titers and RNA levels. Despite the variability we observed for individual samples the overall infectivity differed among the three variants. Both Delta and Epsilon had significantly higher infectivity than Alpha, as measured by the number of infectious units per quantity of viral E gene RNA (6 and 4 times as much, p=0.0002 and 0.009 respectively) or subgenomic E RNA (11 and 7 times as much, p<0.0001 and 0.006 respectively). Conclusion In addition to higher viral RNA levels reported for the Delta variant, the infectivity (amount of replication competent virus per viral genome copy) may also be increased compared to Alpha. Measuring the relationship between live virus and viral RNA is an important step in assessing the infectivity of novel SARS-CoV-2 variants. An increase in the infectivity of the Delta variant may further explain increased spread and suggests a need for increased measures to prevent viral transmission.

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