scholarly journals Sample collection and transport strategies to enhance yield, accessibility, and biosafety of COVID-19 RT-PCR testing

2021 ◽  
Vol 70 (9) ◽  
Author(s):  
Padmapriya Banada ◽  
David Elson ◽  
Naranjargal Daivaa ◽  
Claire Park ◽  
Samuel Desind ◽  
...  
Keyword(s):  
Diagnostic Yield ◽  
Point Of Care ◽  
Sampling Strategy ◽  
Sample Collection ◽  
Rt Pcr ◽  
Non Invasive ◽  
Viral Transport ◽  
Combined Use ◽  
Nasal Swabs ◽  
Polymerase Chain

Introduction. Non-invasive sample collection and viral sterilizing buffers have independently enabled workflows for more widespread COVID-19 testing by reverse-transcriptase polymerase chain reaction (RT-PCR). Gap statement. The combined use of sterilizing buffers across non-invasive sample types to optimize sensitive, accessible, and biosafe sampling methods has not been directly and systematically compared. Aim. We aimed to evaluate diagnostic yield across different non-invasive samples with standard viral transport media (VTM) versus a sterilizing buffer eNAT- (Copan diagnostics Murrieta, CA) in a point-of-care diagnostic assay system. Methods. We prospectively collected 84 sets of nasal swabs, oral swabs, and saliva, from 52 COVID-19 RT-PCR-confirmed patients, and nasopharyngeal (NP) swabs from 37 patients. Nasal swabs, oral swabs, and saliva were placed in either VTM or eNAT, prior to testing with the Xpert Xpress SARS-CoV-2 (Xpert). The sensitivity of each sampling strategy was compared using a composite positive standard. Results. Swab specimens collected in eNAT showed an overall superior sensitivity compared to swabs in VTM (70 % vs 57 %, P=0.0022). Direct saliva 90.5 %, (95 % CI: 82 %, 95 %), followed by NP swabs in VTM and saliva in eNAT, was significantly more sensitive than nasal swabs in VTM (50 %, P<0.001) or eNAT (67.8 %, P=0.0012) and oral swabs in VTM (50 %, P<0.0001) or eNAT (58 %, P<0.0001). Saliva and use of eNAT buffer each increased detection of SARS-CoV-2 with the Xpert; however, no single sample matrix identified all positive cases. Conclusion. Saliva and eNAT sterilizing buffer can enhance safe and sensitive detection of COVID-19 using point-of-care GeneXpert instruments.

scholarly journals Evaluation of sample collection and transport strategies to enhance yield, accessibility, and biosafety of COVID-19 RT-PCR testing

2021 ◽  
Author(s):  
Padmapriya Banada ◽  
David Elson ◽  
Naranjargal Daivaa ◽  
Claire Park ◽  
Samuel Desind ◽  
...  
Keyword(s):  
Sampling Methods ◽  
Diagnostic Yield ◽  
Sampling Strategy ◽  
Sample Collection ◽  
Rt Pcr ◽  
Viral Inactivation ◽  
Viral Transport ◽  
Nasal Swabs ◽  
Polymerase Chain ◽  
Transport Buffer

ABSTRACTSensitive, accessible, and biosafe sampling methods for COVID-19 reverse-transcriptase polymerase chain reaction (RT-PCR) assays are needed for frequent and widespread testing. We systematically evaluated diagnostic yield across different sample collection and transport workflows, including the incorporation of a viral inactivation buffer. We prospectively collected nasal swabs, oral swabs, and saliva, from 52 COVID-19 RT-PCR-confirmed patients, and nasopharyngeal (NP) swabs from 37 patients. Nasal and oral swabs were placed in both viral transport media (VTM) and eNAT™, a sterilizing transport buffer, prior to testing with the Xpert Xpress SARS-CoV-2 (Xpert) test. The sensitivity of each sampling strategy was compared using a composite positive standard. Overall, swab specimens collected in eNAT showed superior sensitivity compared to swabs in VTM (70% vs 57%, P=0.0022). Direct saliva 90.5%, (95% CI: 82%, 95%), followed by NP swabs in VTM and saliva in eNAT, was significantly more sensitive than nasal swabs in VTM (50%, P<0.001) or eNAT (67.8%, P=0.0012) and oral swabs in VTM (50%, P<0.0001) or eNAT (56%, P<0.0001). Saliva and use of eNAT buffer each increased detection of SARS-CoV-2 with the Xpert test; however, no single sample matrix identified all positive cases.

scholarly journals Diagnostic accuracy of a novel SARS-CoV-2 antigen-detecting rapid diagnostic test from standardized self-collected anterior nasal swabs

2021 ◽  
Author(s):  
Bilgin Osmanodja ◽  
Klemens Budde ◽  
Daniel Zickler ◽  
Marcel G. Naik ◽  
Jörg Hofmann ◽  
...  
Keyword(s):  
Primary Endpoint ◽  
Point Of Care ◽  
High Viral Load ◽  
Ease Of Use ◽  
Healthcare Personnel ◽  
Sample Collection ◽  
Rt Pcr ◽  
Self Testing ◽  
Nasal Swabs ◽  
Potential Benefits

AbstractBackgroundAntigen-detecting rapid diagnostic tests (Ag-RDT) for SARS-CoV-2 offer new opportunities for the quick and laboratory-independent identification of infected individuals for control of the SARS-CoV-2 pandemic. Despite the potential benefits, nasopharyngeal sample collection is frequently perceived as uncomfortable by patients and requires trained healthcare personnel with protective equipment. Therefore, anterior nasal self-sampling is increasingly recognized as a valuable alternative.MethodsWe performed a prospective, single-center, point of care validation of an Ag-RDT using a polypropylene absorbent collector for standardized self-collected anterior nasal swabs. Real-Time Polymerase Chain Reaction (RT-PCR) from combined oropharyngeal/nasopharyngeal swabs served as a comparator. Primary endpoint was sensitivity of the standardized Ag-RDT in symptomatic patients with medium or high viral concentration (≥ 1 million RNA copies on RT-PCR for SARS-CoV-2).ResultsBetween February 12 and March 22, 2021, 388 participants were enrolled. After exclusion of 9 patients for which no PCR result could be obtained, the novel Ag-RDT was evaluated based on 379 participants, of which 273 were symptomatic and 106 asymptomatic. In 61 samples from symptomatic patients with medium or high viral load (≥ 1 million RNA copies), the sensitivity of the standardized Ag-RDT was 96.7% (59/61; 95%CI: 88.7-99.6%) for the primary endpoint. In total, 62 positive Ag-RDT results were detected out of 70 RT-PCR positive individuals, yielding an overall sensitivity of 88.6% (95%CI: 78.7-94.9%). Specificity was 99.7% (95%CI: 98.2-100%) in 309 RT-PCR negative individuals.ConclusionHere, we present a validation of a novel Ag-RDT with a standardized sampling process for anterior nasal self-collection, which meets WHO criteria of ≥80% sensitivity and ≥97% specificity. Although less sensitive than RT-PCR, this assay could be beneficial due to its rapid results, ease of use, and suitability for standardized self-testing.(Funded by Drägerwerk AG & Co. KGaA, Lübeck, Germany; ClinicalTrials.gov number NCT04698993)

scholarly journals A smartphone-read ultrasensitive and quantitative saliva test for COVID-19

2020 ◽  
Vol 7 (2) ◽  
pp. eabe3703
Author(s):  
Bo Ning ◽  
Tao Yu ◽  
Shengwei Zhang ◽  
Zhen Huang ◽  
Di Tian ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Rna Isolation ◽  
Rt Pcr ◽  
Laboratory Equipment ◽  
Viral Loads ◽  
Nasal Swabs ◽  
Polymerase Chain ◽  
Potential Use ◽  
Saliva Test

Point-of-care COVID-19 assays that are more sensitive than the current RT-PCR (reverse transcription polymerase chain reaction) gold standard assay are needed to improve disease control efforts. We describe the development of a portable, ultrasensitive saliva-based COVID-19 assay with a 15-min sample-to-answer time that does not require RNA isolation or laboratory equipment. This assay uses CRISPR-Cas12a activity to enhance viral amplicon signal, which is stimulated by the laser diode of a smartphone-based fluorescence microscope device. This device robustly quantified viral load over a broad linear range (1 to 105 copies/μl) and exhibited a limit of detection (0.38 copies/μl) below that of the RT-PCR reference assay. CRISPR-read SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) RNA levels were similar in patient saliva and nasal swabs, and viral loads measured by RT-PCR and the smartphone-read CRISPR assay demonstrated good correlation, supporting the potential use of this portable assay for saliva-based point-of-care COVID-19 diagnosis.

scholarly journals Accuracy of a Novel SARS-CoV-2 Antigen-Detecting Rapid Diagnostic Test from Standardized Self-Collected Anterior Nasal Swabs

2021 ◽  
Vol 10 (10) ◽  
pp. 2099
Author(s):  
Bilgin Osmanodja ◽  
Klemens Budde ◽  
Daniel Zickler ◽  
Marcel G. Naik ◽  
Jörg Hofmann ◽  
...  
Keyword(s):  
Primary Endpoint ◽  
Point Of Care ◽  
World Health Organisation ◽  
Ease Of Use ◽  
Alternative Methods ◽  
World Health ◽  
Healthcare Personnel ◽  
Sample Collection ◽  
Rt Pcr ◽  
Nasal Swabs

Background Antigen-detecting rapid diagnostic tests (Ag-RDT) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) offer new opportunities for the quick and laboratory-independent identification of infected individuals for control of the SARS-CoV-2 pandemic. Despite the potential benefits, nasopharyngeal sample collection is frequently perceived as uncomfortable by patients and requires trained healthcare personnel with protective equipment. Therefore, anterior nasal self-sampling is increasingly recognized as a valuable alternative. Methods We performed a prospective, single-center, point of care validation of an Ag-RDT using a polypropylene absorbent collector for standardized self-collected anterior nasal swabs. Real-time polymerase chain reaction (RT-PCR) from combined oropharyngeal/nasopharyngeal swabs served as a comparator. Primary endpoint was sensitivity of the standardized Ag-RDT in symptomatic patients with medium or high viral concentration (≥1 million RNA copies on RT-PCR for SARS-CoV-2). Results Between 12 February and 22 March 2021, 388 participants were enrolled. After exclusion of 9 patients for which no PCR result could be obtained, the novel Ag-RDT was evaluated based on 379 participants, of whom 273 were symptomatic and 106 asymptomatic. In 61 samples from symptomatic patients with medium or high viral load (≥1 million RNA copies), the sensitivity of the standardized Ag-RDT was 96.7% (59/61; 95% confidence interval (CI): 88.7–99.6%) for the primary endpoint. In total, 62 positive Ag-RDT results were detected out of 70 RT-PCR positive individuals, yielding an overall sensitivity of 88.6% (95% CI: 78.7–94.9%). Specificity was 99.7% (95% CI: 98.2–100%) in 309 RT-PCR negative individuals. Conclusions Here, we present a validation of a novel Ag-RDT with a standardized sampling process for anterior nasal self-collection, which meets World Health Organisation (WHO) criteria of ≥80% sensitivity and ≥97% specificity. Although less sensitive than RT-PCR, this assay could be beneficial due to its rapid results, ease of use, and suitability for standardized self-testing.

Effective optimization of SARS-CoV-2 laboratory testing variables in an era of supply chain constraints

2020 ◽  
Vol 15 (15) ◽  
pp. 1483-1487
Author(s):  
Nikhil S Sahajpal ◽  
Ashis K Mondal ◽  
Allan Njau ◽  
Sudha Ananth ◽  
Kimya Jones ◽  
...  
Keyword(s):  
Supply Chain ◽  
Rna Extraction ◽  
Nasopharyngeal Swab ◽  
Sample Collection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Short Supply ◽  
Viral Transport ◽  
3D Printed ◽  
Bridging Studies

RT-PCR-based assays for the detection of SARS-CoV-2 have played an essential role in the current COVID-19 pandemic. However, the sample collection and test reagents are in short supply, primarily due to supply chain issues. Thus, to eliminate testing constraints, we have optimized three key process variables: RNA extraction and RT-PCR reactions, different sample types and media to facilitate SARS-CoV-2 testing. By performing various validation and bridging studies, we have shown that various sample types such as nasopharyngeal swab, bronchioalveolar lavage and saliva, collected using conventional nasopharyngeal swabs, ESwab or 3D-printed swabs and, preserved in viral transport media, universal transport media, 0.9% sodium chloride or Amies media are compatible with RT-PCR assay for COVID-19. Besides, the reduction of PCR reagents by up to fourfold also produces reliable results.

scholarly journals Screening, Diagnostic and Prognostic Tests for COVID-19: A Comprehensive Review

Life ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 561
Author(s):  
Mariana Ulinici ◽  
Serghei Covantev ◽  
James Wingfield-Digby ◽  
Apostolos Beloukas ◽  
Alexander G. Mathioudakis ◽  
...  
Keyword(s):  
Point Of Care ◽  
Standard Test ◽  
Current Evidence ◽  
Molecular Testing ◽  
Rt Pcr ◽  
Diagnostic Screening ◽  
Comprehensive Review ◽  
Prognostic Tests ◽  
Polymerase Chain ◽  
Antigen Testing

While molecular testing with real-time polymerase chain reaction (RT-PCR) remains the gold-standard test for COVID-19 diagnosis and screening, more rapid or affordable molecular and antigen testing options have been developed. More affordable, point-of-care antigen testing, despite being less sensitive compared to molecular assays, might be preferable for wider screening initiatives. Simple laboratory, imaging and clinical parameters could facilitate prognostication and triage. This comprehensive review summarises current evidence on the diagnostic, screening and prognostic tests for COVID-19.

scholarly journals CT findings of 795 COVID-19 positive cases: a multicenter study in Egypt

2020 ◽  
Vol 51 (1) ◽  
Author(s):  
Youssriah Yahia Sabri ◽  
Mohamed Mohsen Tolba Fawzi ◽  
Eman Zaki Nossair ◽  
Safaa Mohamed El-Mandooh ◽  
Amira Aly Hegazy ◽  
...  
Keyword(s):  
Virus Disease ◽  
Chest Ct ◽  
Imaging Features ◽  
Rt Pcr ◽  
Relevant Research ◽  
Non Invasive ◽  
Ct Findings ◽  
Global Pandemic ◽  
Polymerase Chain ◽  
Ct Studies

Abstract Background Corona Virus Disease 2019 (COVID-19) outbreak was officially announced as a global pandemic by the WHO on March 11th 2020. Thorough understanding of CT imaging features of COVID-19 is essential for effective patient management; rationalizing the need for relevant research. The aim of this study was to analyze the chest CT findings of patients with real-time polymerase chain reaction (RT-PCR) proved COVID-19 admitted to four Egyptian hospitals. The recently published RSNA expert consensus statement on reporting COVID-19 chest CT findings was taken into consideration. Results Normal CT “negative for COVID-19” was reported in 26.1% of our RT-PCR proved COVID-19 cases. In descending order of prevalence, imaging findings of the positive CT studies (73.9%) included GGO (69%), consolidation (49.7%), crazy paving (15.4%), and peri-lobular fibrosis (40.6%). These showed a dominantly bilateral (68.2%), peripheral (72.4%), and patchy (64.7%) distribution. Remarkably, thymic hyperplasia was identified in 14.3% of studies. According to the RSNA consensus, CT findings were classified as typical in 68.9%, indeterminate in 3.6%, and atypical in 1.4% of the evaluated CT studies. Conclusion Although COVID-19 cannot be entirely excluded by chest CT, it can be distinguished in more than two-thirds of cases; making CT a widely available, non-invasive, and rapid diagnostic tool.

scholarly journals Diagnosis of SARS-Cov-2 Infection by RT-PCR Using Specimens Other Than Naso- and Oropharyngeal Swabs: A Systematic Review and Meta-Analysis

Diagnostics ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 363
Author(s):  
Vânia M. Moreira ◽  
Paulo Mascarenhas ◽  
Vanessa Machado ◽  
João Botelho ◽  
José João Mendes ◽  
...  
Keyword(s):  
Systematic Review ◽  
Clinical Performance ◽  
Point Of Care ◽  
Meta Analysis ◽  
High Sensitivity ◽  
Nasopharyngeal Swab ◽  
Sample Collection ◽  
Rt Pcr ◽  
Oropharyngeal Swab ◽  
Nucleic Acid Assays

The rapid and accurate testing of SARS-CoV-2 infection is still crucial to mitigate, and eventually halt, the spread of this disease. Currently, nasopharyngeal swab (NPS) and oropharyngeal swab (OPS) are the recommended standard sampling techniques, yet, these have some limitations such as the complexity of collection. Hence, several other types of specimens that are easier to obtain are being tested as alternatives to nasal/throat swabs in nucleic acid assays for SARS-CoV-2 detection. This study aims to critically appraise and compare the clinical performance of RT-PCR tests using oral saliva, deep-throat saliva/posterior oropharyngeal saliva (DTS/POS), sputum, urine, feces, and tears/conjunctival swab (CS) against standard specimens (NPS, OPS, or a combination of both). In this systematic review and meta-analysis, five databases (PubMed, Scopus, Web of Science, ClinicalTrial.gov and NIPH Clinical Trial) were searched up to the 30th of December, 2020. Case-control and cohort studies on the detection of SARS-CoV-2 were included. The methodological quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS 2). We identified 1560 entries, 33 of which (1.1%) met all required criteria and were included for the quantitative data analysis. Saliva presented the higher accuracy, 92.1% (95% CI: 70.0–98.3), with an estimated sensitivity of 83.9% (95% CI: 77.4–88.8) and specificity of 96.4% (95% CI: 89.5–98.8). DTS/POS samples had an overall accuracy of 79.7% (95% CI: 43.3–95.3), with an estimated sensitivity of 90.1% (95% CI: 83.3–96.9) and specificity of 63.1% (95% CI: 36.8–89.3). The remaining index specimens could not be adequately assessed given the lack of studies available. Our meta-analysis shows that saliva samples from the oral region provide a high sensitivity and specificity; therefore, these appear to be the best candidates for alternative specimens to NPS/OPS in SARS-CoV-2 detection, with suitable protocols for swab-free sample collection to be determined and validated in the future. The distinction between oral and extra-oral salivary samples will be crucial, since DTS/POS samples may induce a higher rate of false positives. Urine, feces, tears/CS and sputum seem unreliable for diagnosis. Saliva testing may increase testing capacity, ultimately promoting the implementation of truly deployable COVID-19 tests, which could either work at the point-of-care (e.g. hospitals, clinics) or at outbreak control spots (e.g., schools, airports, and nursing homes).

scholarly journals Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252687
Author(s):  
Sukalyani Banik ◽  
Kaheerman Saibire ◽  
Shraddha Suryavanshi ◽  
Glenn Johns ◽  
Soumitesh Chakravorty ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Viral Rna ◽  
Clinical Samples ◽  
Sample Collection ◽  
Rt Pcr ◽  
Diagnostic Assays ◽  
Guanidinium Thiocyanate ◽  
Upper Respiratory ◽  
Polymerase Chain ◽  
The Stability

Background Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings. Methods We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C. Results SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log10 PFU/ml within a minute of incubation. When saliva was diluted 1:1 in eNAT, no cytopathic effect (CPE) on VeroE6 cells was observed, although SARS-CoV-2 RNA could be detected even after 30 min incubation and after two cell culture passages. A 1:2 (saliva:eNAT) dilution abrogated both CPE and detectable viral RNA after as little as 5 min incubation in eNAT. SARS-CoV-2 RNA from virus spiked at 5X the limit of detection remained positive up to 7 days of incubation in all tested conditions. Conclusion eNAT and similar guanidinium thiocyanate-based media may be of value for transport, stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection.

scholarly journals Point-of-Care Diagnostics of COVID-19: From Current Work to Future Perspectives

Sensors ◽  
2020 ◽  
Vol 20 (15) ◽  
pp. 4289 ◽  
Author(s):  
Heba A. Hussein ◽  
Rabeay Y. A. Hassan ◽  
Marco Chino ◽  
Ferdinando Febbraio
Keyword(s):  
Point Of Care ◽  
Virus Detection ◽  
Diagnostic Tools ◽  
Sample Treatment ◽  
Rt Pcr ◽  
Point Of Care Diagnostics ◽  
Electrochemical Immunosensors ◽  
Global Concern ◽  
Polymerase Chain ◽  
Reliable Methods

Coronaviruses have received global concern since 2003, when an outbreak caused by SARS-CoV emerged in China. Later on, in 2012, the Middle-East respiratory syndrome spread in Saudi Arabia, caused by MERS-CoV. Currently, the global crisis is caused by the pandemic SARS-CoV-2, which belongs to the same lineage of SARS-CoV. In response to the urgent need of diagnostic tools, several lab-based and biosensing techniques have been proposed so far. Five main areas have been individuated and discussed in terms of their strengths and weaknesses. The cell-culture detection and the microneutralization tests are still considered highly reliable methods. The genetic screening, featuring the well-established Real-time polymerase chain reaction (RT-PCR), represents the gold standard for virus detection in nasopharyngeal swabs. On the other side, immunoassays were developed, either by screening/antigen recognition of IgM/IgG or by detecting the whole virus, in blood and sera. Next, proteomic mass-spectrometry (MS)-based methodologies have also been proposed for the analysis of swab samples. Finally, virus-biosensing devices were efficiently designed. Both electrochemical immunosensors and eye-based technologies have been described, showing detection times lower than 10 min after swab introduction. Alternative to swab-based techniques, lateral flow point-of-care immunoassays are already commercially available for the analysis of blood samples. Such biosensing devices hold the advantage of being portable for on-site testing in hospitals, airports, and hotspots, virtually without any sample treatment or complicated lab precautions.

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