scholarly journals Pooled sputum to optimise the efficiency and utility of rapid, point-of-care molecular SARS-CoV-2 testing

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Alison Burdett ◽  
Christofer Toumazou ◽  
Rashmita Sahoo ◽  
Adam Mujan ◽  
Tsz-Kin Hon ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
High Sensitivity ◽  
Pool Size ◽  
Pooled Testing ◽  
Paired Samples ◽  
Absolute Strength ◽  
Potential Benefits ◽  
First Time ◽  
Multiple Patients

Abstract Background As SARS-CoV-2 testing expands, particularly to widespread asymptomatic testing, high sensitivity point-of-care PCR platforms may optimise potential benefits from pooling multiple patients’ samples. Method We tested patients and asymptomatic citizens for SARS-CoV-2, exploring the efficiency and utility of CovidNudge (i) for detection in individuals’ sputum (compared to nasopharyngeal swabs), (ii) for detection in pooled sputum samples, and (iii) by modelling roll out scenarios for pooled sputum testing. Results Across 295 paired samples, we find no difference (p = 0.1236) in signal strength for sputum (mean amplified replicates (MAR) 25.2, standard deviation (SD) 14.2, range 0–60) compared to nasopharyngeal swabs (MAR 27.8, SD 12.4, range 6–56). At 10-sample pool size we find some drop in absolute strength of signal (individual sputum MAR 42.1, SD 11.8, range 13–60 vs. pooled sputum MAR 25.3, SD 14.6, range 1–54; p < 0.0001), but only marginal drop in sensitivity (51/53,96%). We determine a limit of detection of 250 copies/ml for an individual test, rising only four-fold to 1000copies/ml for a 10-sample pool. We find optimal pooled testing efficiency to be a 12–3-1-sample model, yet as prevalence increases, pool size should decrease; at 5% prevalence to maintain a 75% probability of negative first test, 5-sample pools are optimal. Conclusion We describe for the first time the use of sequentially dipped sputum samples for rapid pooled point of care SARS-CoV-2 PCR testing. The potential to screen asymptomatic cohorts rapidly, at the point-of-care, with PCR, offers the potential to quickly identify and isolate positive individuals within a population “bubble”.

scholarly journals Ag nanoparticles outperform Au nanoparticles for the use as label in electrochemical point-of-care sensors

2021 ◽  
Author(s):  
Franziska Beck ◽  
Carina Horn ◽  
Antje J. Baeumner
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Automatic Measurement ◽  
High Sensitivity ◽  
Differential Pulse ◽  
Small Sample ◽  
Blood Protein ◽  
User Friendliness ◽  
Response Curves ◽  
Assay Protocol

AbstractElectrochemical immunosensors enable rapid analyte quantification in small sample volumes, and have been demonstrated to provide high sensitivity and selectivity, simple miniaturization, and easy sensor production strategies. As a point-of-care (POC) format, user-friendliness is equally important and most often not combinable with high sensitivity. As such, we demonstrate here that a sequence of metal oxidation and reduction, followed by stripping via differential pulse voltammetry (DPV), provides lowest limits of detection within a 2-min automatic measurement. In exchanging gold nanoparticles (AuNPs), which dominate in the development of POC sensors, with silver nanoparticles (AgNPs), not only better sensitivity was obtained, but more importantly, the assay protocol could be simplified to match POC requirements. Specifically, we studied both nanoparticles as reporter labels in a sandwich immunoassay with the blood protein biomarker NT-proBNP. For both kinds of nanoparticles, the dose-response curves easily covered the ng∙mL−1 range. The mean standard deviation of all measurements of 17% (n ≥ 4) and a limit of detection of 26 ng∙mL−1 were achieved using AuNPs, but their detection requires addition of HCl, which is impossible in a POC format. In contrast, since AgNPs are electrochemically less stable, they enabled a simplified assay protocol and provided even lower LODs of 4.0 ng∙mL−1 in buffer and 4.7 ng∙mL−1 in human serum while maintaining the same or even better assay reliability, storage stability, and easy antibody immobilization protocols. Thus, in direct comparison, AgNPs clearly outperform AuNPs in desirable POC electrochemical assays and should gain much more attention in the future development of such biosensors.

scholarly journals Direct-RT-qPCR Detection of SARS-CoV-2 without RNA Extraction as Part of a COVID-19 Testing Strategy: From Sample to Result in One Hour

Diagnostics ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 605 ◽  
Author(s):  
Eva Kriegova ◽  
Regina Fillerova ◽  
Petr Kvapil
Keyword(s):  
High Throughput Screening ◽  
High Speed ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Rna Extraction ◽  
Rna Isolation ◽  
High Sensitivity ◽  
Ease Of Use ◽  
Diagnostic Tools ◽  
Heat Inactivation

Due to the lack of protective immunity in the general population and the absence of effective antivirals and vaccines, the Coronavirus disease 2019 (COVID-19) pandemic continues in some countries, with local epicentres emerging in others. Due to the great demand for effective COVID-19 testing programmes to control the spread of the disease, we have suggested such a testing programme that includes a rapid RT-qPCR approach without RNA extraction. The Direct-One-Step-RT-qPCR (DIOS-RT-qPCR) assay detects severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in less than one hour while maintaining the high sensitivity and specificity required of diagnostic tools. This optimised protocol allows for the direct use of swab transfer media (14 μL) without the need for RNA extraction, achieving comparable sensitivity to the standard method that requires the time-consuming and costly step of RNA isolation. The limit of detection for DIOS-RT-qPCR was lower than seven copies/reaction, which translates to 550 virus copies/mL of swab. The speed, ease of use and low price of this assay make it suitable for high-throughput screening programmes. The use of fast enzymes allows RT-qPCR to be performed under standard laboratory conditions within one hour, making it a potential point-of-care solution on high-speed cycling instruments. This protocol also implements the heat inactivation of SARS-CoV-2 (75 °C for 10 min), which renders samples non-infectious, enabling testing in BSL-2 facilities. Moreover, we discuss the critical steps involved in developing tests for the rapid detection of COVID-19. Implementing rapid, easy, cost-effective methods can help control the worldwide spread of the COVID-19 infection.

scholarly journals Multiplex Detection of Three Select Agents Directly from Blood by Use of the GeneXpert System

2019 ◽  
Vol 57 (5) ◽  
Author(s):  
Padmapriya P. Banada ◽  
Srinidhi Deshpande ◽  
Sukalyani Banik ◽  
Darshini Shah ◽  
Ranie Koshy ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Severe Disease ◽  
High Sensitivity ◽  
Negative Control ◽  
Blood Samples ◽  
Content Type ◽  
Clinical Patient

ABSTRACT Francisella tularensis, Bacillus anthracis, and Yersinia pestis are tier 1 select agents with the potential to rapidly cause severe disease. Rapid detection of these bacteria from patient samples at the point of care could contribute to improved clinical outcomes in the event of a bioterrorism attack. A multiplex nested PCR assay for detection of F. tularensis, B. anthracis, and Y. pestis directly from patient blood samples was developed using the GeneXpert system. The multiplex GeneXpert cartridge-based assay includes all necessary sample processing and amplification reagents. Blood samples spiked with different numbers of CFU were used to measure the analytical limit of detection (LOD) and dynamic range. Sensitivity was determined by testing spiked blood samples and negative-control blood in a blind manner. Specificity was determined by testing against nontarget pathogens and blood samples from clinical patients. The assay LOD was 8.5 CFU/ml for F. tularensis, 10 CFU/ml for B. anthracis, and 4.5 CFU/ml for Y. pestis. The sensitivity was 100% at the LOD for all three select agent bacteria in spiked patient blood samples. The assay specificity was 100% when it was tested against both nontarget pathogens and clinical patient blood samples. The total assay time was approximately 100 min. This automated assay, which is suitable for use at the point of care, identifies three select agents directly in blood without the need for enrichment with a high sensitivity within 100 min. This assay may enable rapid detection and treatment of patients infected with the target organisms in the event of a bioterrorism attack.

scholarly journals Recent Advances of Field-Effect Transistor Technology for Infectious Diseases

Biosensors ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 103
Author(s):  
Abbas Panahi ◽  
Deniz Sadighbayan ◽  
Saghi Forouhi ◽  
Ebrahim Ghafar-Zadeh
Keyword(s):  
Infectious Diseases ◽  
Field Effect ◽  
Field Effect Transistor ◽  
Limit Of Detection ◽  
Point Of Care ◽  
High Sensitivity ◽  
Cmos Technology ◽  
Oxide Semiconductor ◽  
Label Free ◽  
Effect Transistor

Field-effect transistor (FET) biosensors have been intensively researched toward label-free biomolecule sensing for different disease screening applications. High sensitivity, incredible miniaturization capability, promising extremely low minimum limit of detection (LoD) at the molecular level, integration with complementary metal oxide semiconductor (CMOS) technology and last but not least label-free operation were amongst the predominant motives for highlighting these sensors in the biosensor community. Although there are various diseases targeted by FET sensors for detection, infectious diseases are still the most demanding sector that needs higher precision in detection and integration for the realization of the diagnosis at the point of care (PoC). The COVID-19 pandemic, nevertheless, was an example of the escalated situation in terms of worldwide desperate need for fast, specific and reliable home test PoC devices for the timely screening of huge numbers of people to restrict the disease from further spread. This need spawned a wave of innovative approaches for early detection of COVID-19 antibodies in human swab or blood amongst which the FET biosensing gained much more attention due to their extraordinary LoD down to femtomolar (fM) with the comparatively faster response time. As the FET sensors are promising novel PoC devices with application in early diagnosis of various diseases and especially infectious diseases, in this research, we have reviewed the recent progress on developing FET sensors for infectious diseases diagnosis accompanied with a thorough discussion on the structure of Chem/BioFET sensors and the readout circuitry for output signal processing. This approach would help engineers and biologists to gain enough knowledge to initiate their design for accelerated innovations in response to the need for more efficient management of infectious diseases like COVID-19.

scholarly journals Pooled Sample Testing for SARS-CoV-2 Using Rapid RT-PCR COVID-19 Tests

2021 ◽  
Author(s):  
Bethany Hyde ◽  
Prat Verma ◽  
Ethan M. Berke
Keyword(s):  
Lower Cost ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Rapid Testing ◽  
Rt Pcr ◽  
Pooled Testing ◽  
Testing Performance ◽  
Sample Testing ◽  
Pooled Sample ◽  
Pool Sizes

AbstractWe tested an operationally efficient way to pool samples on a rapid, point-of-care PCR device and examined the limit of detection of SARS-CoV-2 for various pool sizes. Pooled testing maintained testing performance similar to individual sample PCR testing, offering the potential for scalable rapid testing at lower cost with less supplies.

scholarly journals Recommendations for sample pooling on the Cepheid GeneXpert® system using the Cepheid Xpert® Xpress SARS-CoV-2 assay

2020 ◽  
Author(s):  
Michael G. Becker ◽  
Tracy Taylor ◽  
Sandra Kiazyk ◽  
Dana R. Cabiles ◽  
Adrienne F.A. Meyers ◽  
...  
Keyword(s):  
Public Health ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Test Site ◽  
Viral Loads ◽  
Pooled Testing ◽  
Point Of Care Test ◽  
Health Authorities ◽  
Public Health Authorities ◽  
Sample Pooling

AbstractThe coronavirus disease 2019 (Covid-19) pandemic, caused by SARS-CoV-2, has resulted in a global testing supply shortage. In response, pooled testing has emerged as a promising strategy that can immediately increase testing capacity. Here, we provide support for the adoption of sample pooling with the point-of-care Cepheid Xpert® Xpress SARS-CoV-2 molecular assay. Corroborating previous findings, the Xpert® Xpress SARS-CoV-2 assay limit of detection was comparable to central laboratory reverse-transcription quantitative PCR tests with observed SARS-CoV-2 detection below 100 copies/mL. The Xpert® Xpress assay detected SARS-CoV-2 after samples with minimum viral loads of 461 copies/mL were diluted into six sample pools. Based on these data, we recommend the adoption of pooled testing with the Xpert® Xpress SARS-CoV-2 assay where warranted by population public health needs. The suggested number of samples per pool, or pooling depth, is unique for each point-of-care test site and should be determined by assessing positive test rates. To statistically determine appropriate pooling depth, we have calculated the pooling efficiency for numerous combinations of pool sizes and test rates. This information is included as a supplemental dataset that we encourage public health authorities to use as a guide to make recommendations that will maximize testing capacity and resource conservation.

scholarly journals Recommendations for sample pooling on the Cepheid GeneXpert® system using the Cepheid Xpert® Xpress SARS-CoV-2 assay

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0241959
Author(s):  
Michael G. Becker ◽  
Tracy Taylor ◽  
Sandra Kiazyk ◽  
Dana R. Cabiles ◽  
Adrienne F. A. Meyers ◽  
...  
Keyword(s):  
Public Health ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Viral Loads ◽  
Pooled Testing ◽  
Health Authorities ◽  
Public Health Authorities ◽  
Sample Pooling ◽  
Testing Site ◽  
The Individual

The coronavirus disease 2019 (Covid-19) pandemic, caused by SARS-CoV-2, has resulted in a global testing supply shortage. In response, pooled testing has emerged as a promising strategy that can immediately increase testing capacity. In pooled sample testing, multiple samples are combined (or pooled) together and tested as a single unit. If the pool is positive, the individual samples can then be individually tested to identify the positive case(s). Here, we provide support for the adoption of sample pooling with the point-of-care Cepheid Xpert® Xpress SARS-CoV-2 molecular assay. Corroborating previous findings, the limit of detection of this assay was comparable to laboratory-developed reverse-transcription quantitative PCR SARS-CoV-2 tests, with observed detection below 100 copies/mL. The Xpert® Xpress assay detected SARS-CoV-2 after samples with minimum viral loads of 461 copies/mL were pooled in groups of six. Based on these data, we recommend the adoption of pooled testing with the Xpert® Xpress SARS-CoV-2 assay where warranted based on public health needs. The suggested number of samples per pool, or the pooling depth, is unique for each point-of-care testing site and can be determined by the positive test rates. To statistically determine appropriate pooling depth, we have calculated the pooling efficiency for numerous combinations of pool sizes and test rates. This information is included as a supplemental dataset that we encourage public health authorities to use as a guide to make recommendations that will maximize testing capacity and resource conservation.

Determination of sex-specific 99th percentile upper reference limits for a point of care high sensitivity cardiac troponin I assay

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Fred S. Apple ◽  
Karen Schulz ◽  
Christian W. Schmidt ◽  
Trees S. Y. van Domburg ◽  
Judith M. Fonville ◽  
...  
Keyword(s):  
Cardiac Troponin ◽  
Troponin I ◽  
Limit Of Detection ◽  
Point Of Care ◽  
High Sensitivity ◽  
The Novel ◽  
Limit Of Quantitation ◽  
Reference Limits ◽  
Males And Females

Abstract Objectives High sensitivity (hs) cardiac troponin (cTn) assays are defined per the IFCC Committee on Clinical Application of Cardiac Biomarker (C-CB) by the ability to measure ≥ 50% of concentrations greater than the limit of detection (LoD) with an impression of ≤10% at sex-specific 99th percentiles. Our study determined the sex-specific 99th percentile upper reference limits for males and females utilizing heparinized plasma from AACC universal sample bank for the Siemens point of care (POC) Atellica® VTLi hs-cTnI immunoassay. Methods Apparently healthy subjects, included overall 693, males 363, and females 330, following exclusionary surrogate biomarker use of hemoglobin A1c, NT-proBNP, and eGFR, along with statin medication. hs-cTnI was measured in a central laboratory, on multiple POC Atellica® VTLi immunoassay analyzers. The LoD was 1.24 ng/L and limit of quantitation (CV 20%) was 6.7 ng/L. 99th percentile URLs were determined by the nonparametric (NP) method. Results Histograms of the hs-cTnI concentrations (ng/L) for males and females were used to visualize the distributions and concentrations in men and women and differed significantly (pre- and post-exclusion, both p <0.001). 99th percentile URLs were: overall 23 ng/L (90% CI 20–32 ng/L); male 27 ng/L (CI 21–37 ng/L); female 18 ng/L (CI 9–78 ng/L). The percentages of subjects having a measurable concentration ≥ the LoD were: overall 83.7%, male 87.3%, female 79.7%. Conclusions Our findings show the novel POC Atellica® VTLi hs-cTnI assay meets the designation of a ‘high-sensitivity’ assay using heparinized plasma.

scholarly journals DEVELOPMENT OF ARGININE DEIMINASE BASED CONDUCTOMETRIC BIOSENSOR FOR ARGININE DETERMINATION

2021 ◽  
Vol 18 (2) ◽  
pp. 4-13
Author(s):  
O. O. Soldatkin ◽  
I. S. Kucherenko ◽  
O. Ya. Sayapina ◽  
D. Yu. Kucherenko ◽  
S. V. Marchenko ◽  
...  
Keyword(s):  
Ionic Strength ◽  
Buffer Capacity ◽  
Limit Of Detection ◽  
High Sensitivity ◽  
Arginine Deiminase ◽  
Real Samples ◽  
Covalent Crosslinking ◽  
Enzyme Biosensor ◽  
First Time ◽  
Conductometric Biosensor

For the first time, a conductometric enzyme biosensor was developed to determine arginine concentrations. The bioselective membrane of the biosensor was formed by immobilization of arginine deiminase on the surface of gold planar transducer using covalent crosslinking of glutaraldehyde with bovine serum albumin. An effect of the solution characteristics (ionic strength, buffer capacity) on the biosensor functioning was studied. The proposed monoenzyme biosensor was shown to have high sensitivity to arginine (minimum limit of detection - 5 μM) and good selectivity towards possible interferents. The linear range of determination was from 10 to 800 μM. The biosensor sensitivity to arginine is 72 μS /μM. The developed biosensor was demonstrated to be promising for the arginine analysis in real samples.

Giant Magnetoresistive Based Handheld System for Rapid Detection of Human NT-proBNP

2019 ◽  
Author(s):  
Wei Wang ◽  
Todd Klein ◽  
James Collins
Keyword(s):  
Limit Of Detection ◽  
Detection System ◽  
Point Of Care ◽  
High Sensitivity ◽  
Time Signal ◽  
Point Of Care Testing ◽  
Detection Range ◽  
Further Development ◽  
Signal Readout

In this work, we developed giant magnetoresistive (GMR) based handheld biosensing systems that serve as platform for detecting human NT-proBNP. This assay takes advantages of high sensitivity and real-time signal readout of GMR biosensor. The limit of detection was estimated to be less than 0.01ng/mL, and detection range covered from 0.01 ng/mL to 5 ng/mL was obtained. The assay can be completed within 20 min, which is very important for further development of point-of-care testing. The proposed GMR handheld system is also successfully used for the detection of real NT-proBNP human samples. It can be foreseen that this handheld detection system could become a robust contender in the applications of in vitro biomarker diagnostics.

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