scholarly journals Clinical Evaluation of the Novel Rapid Nucleic Acid Amplification Point-of-Care Test (Smart Gene SARS-CoV-2) in the analysis of Nasopharyngeal and Anterior Nasal samples.

2021 ◽  
Author(s):  
Yoshihiko Kiyasu ◽  
Masato Owaku ◽  
Yusaku Akashi ◽  
Yuto Takeuchi ◽  
Kenji Narahara ◽  
...  
Keyword(s):  
Real Time ◽  
Clinical Performance ◽  
Point Of Care ◽  
Nucleic Acid Amplification ◽  
Molecular Testing ◽  
The Novel ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Point Of Care Test ◽  
Hands On

Introduction Smart Gene is a point-of-care (POC)-type automated molecular testing platform that can be performed with 1 minute of hands-on-time. Smart Gene SARS-CoV-2 is a newly developed Smart Gene molecular assay for the detection of SARS-CoV-2. The analytical and clinical performance of Smart Gene SARS-CoV-2 has not been evaluated. Methods Nasopharyngeal and anterior nasal samples were prospectively collected from subjects referred to the local PCR center from March 25 to July 5, 2021. Two swabs were simultaneously obtained for the Smart Gene SARS-CoV-2 assay and the reference real-time RT-PCR assay, and the results of Smart Gene SARS-CoV-2 were compared to the reference real-time RT-PCR assay. Results Among a total of 1150 samples, 68 of 791 nasopharyngeal samples and 51 of 359 anterior nasal samples were positive for SARS-CoV-2 in the reference real-time RT-PCR assay. In the testing of nasopharyngeal samples, Smart Gene SARS-CoV-2 showed the total, positive and negative concordance of 99.2% (95% confidence interval [CI]: 98.4–99.7%), 94.1% (95% CI: 85.6–98.4%) and 99.7% (95% CI: 99.0–100%), respectively. For anterior nasal samples, Smart Gene SARS-CoV-2 showed the total, positive and negative concordance of 98.9% (95% CI: 97.2–99.7%), 98.0% (95% CI: 89.6–100%) and 99.0% (95% CI: 97.2–99.8%), respectively. In total, 5 samples were positive in the reference real-time RT-PCR and negative in Smart Gene SARS-CoV-2, whereas 5 samples were negative in the reference real-time RT-PCR and positive in Smart Gene SARS-CoV-2. Conclusion Smart Gene SARS-CoV-2 showed sufficient analytical performance for the detection of SARS-CoV-2 in nasopharyngeal and anterior nasal samples.

scholarly journals Comparative Study of the Efficacy of the Abbott ID Now Rapid Assay with Real Time PCR for the Diagnosis of SARS-CoV-2 RNA

2021 ◽  
Author(s):  
Pratiksha Chheda ◽  
Dama Tavisha ◽  
Bhalerao Rahul ◽  
Bagwan Jamir ◽  
Bhat Devdatta ◽  
...  
Keyword(s):  
Real Time ◽  
Point Of Care ◽  
Turnaround Time ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Clinical Patient ◽  
Point Of Care Test ◽  
Percent Agreement ◽  
Current Outbreak ◽  
Spread Of Infection

Abstract Rapid diagnostic tests are of great importance in hospital settings during the current outbreak of SARS-CoV-2. The clinical patient management and spread of infection is critically dependent on molecular assays with shortest possible turn-around time. Here we report performance of a point of care Abbott ID NOW COVID-19 assay in comparison to routinely used real-time RT-PCR assay on 205 clinical specimens. Overall agreement of ID NOW was found to be 93.7% with positive percent agreement (PPA) of 91.8% and negative percent agreement (NPA) of 95.4%. Based on our findings, low turnaround time, minimal infrastructure need and ease of performing the assay, Abbott ID NOW COVID-19 assay can be considered as a point of care test in hospital settings.

Multiplex SARS-CoV-2 RT-qPCR protocol v1

2021 ◽  
Author(s):  
Peter H L Krijger ◽  
Tim A Hoek ◽  
Sanne Boersma ◽  
Lieke I P M Donders ◽  
Maaike M C Broeders ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Hepatitis C ◽  
Real Time ◽  
Internal Control ◽  
Point Of Care ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Reverse Transcription Pcr ◽  
Point Of Care Test ◽  
Novel Coronavirus

An in-house multiplex RT-qPCR , targeting SARS-CoV-2and PDV as internal control [1][2], developed on QuantStudio 7 Pro Real-Time PCR Systems using Life Technologies Taqman FastVirus 1-step mastermix with E-gene primers and probe as described by Corman et al. and N1 primers and probes as described by the CDC[3, 4]. 1.Clancy, A. eta al., The development of a qualitative real-time RT-PCR assay for the detection of hepatitis C virus European Journal Microbial Infectious Diseases, 2008. 276(12): p.1177. 2.Wolters, F., et al., Multi-center evaluation of cepheid xpert® xpress SARS-CoV-2 point-of-care test during the SARS-CoV-2 pandemic. Journal of Clinical Virology, 2020. 128: p. 104426 3.Corman, V.M., et al., Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill, 2020. 25(3). 4.Lu, X., et al., US CDC Real-Time Reverse Transcription PCR Panel for Detection of Severe Acute Respiratory Syndrome Coronavirus 2.Emerging Infectious Disease journal, 2020. 26(8): p. 1654.

scholarly journals Performing point-of-care molecular testing for SARS-CoV-2 with RNA extraction and isothermal amplification

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0243712
Author(s):  
Pierre Garneret ◽  
Etienne Coz ◽  
Elian Martin ◽  
Jean-Claude Manuguerra ◽  
Elodie Brient-Litzler ◽  
...  
Keyword(s):  
Real Time ◽  
Limit Of Detection ◽  
Clinical Sample ◽  
Point Of Care ◽  
Rna Extraction ◽  
Temperature Cycling ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Molecular Testing ◽  
Rt Pcr

To respond to the urgent need for COVID-19 testing, countries perform nucleic acid amplification tests (NAAT) for the detection of SARS-CoV-2 in centralized laboratories. Real-time RT—PCR (Reverse transcription—Polymerase Chain Reaction), used to amplify and detect the viral RNA., is considered, as the current gold standard for diagnostics. It is an efficient process, but the complex engineering required for automated RNA extraction and temperature cycling makes it incompatible for use in point of care settings [1]. In the present work, by harnessing progress made in the past two decades in isothermal amplification and paper microfluidics, we created a portable test, in which SARS-CoV-2 RNA is extracted, amplified isothermally by RT—LAMP (Loop-mediated Isothermal Amplification), and detected using intercalating dyes or fluorescent probes. Depending on the viral load in the tested samples, the detection takes between twenty minutes and one hour. Using a set of 16 pools of naso-pharyngal swab eluates, we estimated a limit of detection comparable to real-time RT-PCR (i.e. 1 genome copies per microliter of clinical sample) and no cross‐reaction with eight major respiratory viruses currently circulating in Europe. We designed and fabricated an easy-to-use portable device called “COVIDISC” to carry out the test at the point of care. The low cost of the materials along with the absence of complex equipment will expedite the widespread dissemination of this device. What is proposed here is a new efficient tool to help managing the pandemics.

scholarly journals Molecular Detection of a Novel Human Influenza (H1N1) of Pandemic Potential by Conventional and Real-Time Quantitative RT-PCR Assays

2009 ◽  
Vol 55 (8) ◽  
pp. 1555-1558 ◽  
Author(s):  
Leo L M Poon ◽  
K H Chan ◽  
G J Smith ◽  
C S W Leung ◽  
Y Guan ◽  
...  
Keyword(s):  
Real Time ◽  
Influenza A ◽  
H1n1 Virus ◽  
Influenza Viruses ◽  
The Novel ◽  
Human Influenza ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Recent Emergence ◽  
Human H5n1

Abstract Background: Influenza A viruses are medically important viral pathogens that cause significant mortality and morbidity throughout the world. The recent emergence of a novel human influenza A virus (H1N1) poses a serious health threat. Molecular tests for rapid detection of this virus are urgently needed. Methods: We developed a conventional 1-step RT-PCR assay and a 1-step quantitative real-time RT-PCR assay to detect the novel H1N1 virus, but not the seasonal H1N1 viruses. We also developed an additional real-time RT-PCR that can discriminate the novel H1N1 from other swine and human H1 subtype viruses. Results: All of the assays had detection limits for the positive control in the range of 1.0 × 10−4 to 2.0 × 10−3 of the median tissue culture infective dose. Assay specificities were high, and for the conventional and real-time assays, all negative control samples were negative, including 7 human seasonal H1N1 viruses, 1 human H2N2 virus, 2 human seasonal H3N2 viruses, 1 human H5N1 virus, 7 avian influenza viruses (HA subtypes 4, 5, 7, 8, 9, and 10), and 48 nasopharyngeal aspirates (NPAs) from patients with noninfluenza respiratory diseases; for the assay that discriminates the novel H1N1 from other swine and human H1 subtype viruses, all negative controls were also negative, including 20 control NPAs, 2 seasonal human H1N1 viruses, 2 seasonal human H3N2 viruses, and 2 human H5N1 viruses. Conclusions: These assays appear useful for the rapid diagnosis of cases with the novel H1N1 virus, thereby allowing better pandemic preparedness.

scholarly journals The QuantuMDx Q-POC SARS-CoV-2 RT-PCR assay for rapid detection of COVID-19 at point-of-care: preliminary evaluation of a novel technology

2021 ◽  
Author(s):  
Jessica Caffry ◽  
Matthew Selby ◽  
Katie Barr ◽  
George Morgan ◽  
David McGurk ◽  
...  
Keyword(s):  
Point Of Care ◽  
Reference Laboratory ◽  
Preliminary Evaluation ◽  
Rt Pcr ◽  
Reference Test ◽  
Pcr Assay ◽  
Current Standard ◽  
Primary Analysis ◽  
Point Of Care Test ◽  
Control And Management

Background: Accurate, affordable, and rapid point-of-care (PoC) diagnostics are critical to the global control and management of the COVID-19 pandemic. The current standard for accurate diagnosis of SARS-CoV-2 is laboratory-based reverse transcription polymerase chain reaction (RT-PCR). Here, we report a preliminary prospective performance evaluation of the QuantuMDx Q-POC SARS CoV-2 RT-PCR assay. Methods: Between November 2020 and March 2021, we obtained 49 longitudinal nose and throat swabs from 29 individuals hospitalised with RT-PCR confirmed COVID-19 at St Georges' NHS Foundation Trust, London (UK). In addition, we obtained 101 mid nasal swabs from healthy volunteers in June 2021. We then used these samples to evaluate the Q-POC SARS-CoV-2 RT-PCR assay. The primary analysis was to compare the sensitivity and specificity of the Q-POC test against a reference laboratory-based RT-PCR assay. Results: The overall sensitivity of the Q-POC test compared with the reference test was 96.88% (83.78%- 99.92% CI) for a cycle threshold (Ct) cut-off value for the reference test of 35 and 80.00% (64.35% to 90.95% CI) without altering the reference test's Ct cut-off value of 40. Conclusions: The Q-POC test is a sensitive, specific and rapid point-of-care test for SARS-CoV-2 at a reference Ct cut-off value of 35. The Q-POC test provides an accurate and afforda-ble option for RT-PCR at point-of-care without the need for sample pre-processing and laboratory handling. The Q-POC test would enable rapid diagnosis and clinical triage in acute care and other settings.

scholarly journals Evaluating Performance of Multiplex Real Time PCR for the Diagnosis of Malaria at Elimination Targeted Low Transmission Settings of Ethiopia.

2021 ◽  
Author(s):  
Mahlet Belachew ◽  
Mistire Wolde ◽  
Desalegn Nega ◽  
Bokretsion Gidey ◽  
Legessie Negash ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Real Time ◽  
Malaria Elimination ◽  
Real Time Pcr ◽  
Nucleic Acid Amplification ◽  
Molecular Testing ◽  
Pcr Assay ◽  
Blood Samples ◽  
Low Transmission ◽  
Highly Sensitive

Abstract Background: Malaria incidence has declined in Ethiopia in the past ten years. Current malaria diagnostic tests, including light microscopy and antigen-detecting rapid tests (RDTs) cannot reliably detect low-density infections. Studies have shown that nucleic acid amplification tests are highly sensitive and specific in detecting malaria infection. Thus, this study took place with the aim of evaluating the performance of multiplex real time PCR for the diagnosis of malaria using patient samples collected from health facilities located at malaria elimination targeted low transmission settings in Ethiopia. Methods: A health facility based cross sectional survey was conducted in selected malaria sentinel sites. Malaria suspected febrile outpatients referred to laboratory for malaria testing between December 2019 and March 2020 were enrolled into this study. Socio demographic information and capillary blood samples were collected from the study participants and tested at spot with RDTs. Additionally, five circles of dry blood sample (DBS) samples on Whatman filter paper and thick and thin smear were prepared for molecular testing and microscopic examination, respectively. Multiplex real time PCR assay was performed at EPHI malaria laboratory. The performance of multiplex real time PCR assay, microscopy and RDT for the diagnosis of malaria was compared and evaluated against each other.Results: Out of 271 blood samples, multiplex real time PCR identified 69 malaria cases as P. falciparum infection, 16 as P. vivax and 3 as mixed infections. Of the total samples, light microscopy detected 33 as Pf, 18 as PV and RDT detected 43 as Pf, 17 as PV, and one mixed infection. Using light microscopy as reference test, the sensitivity and specificity of multiplex real time PCR were 100% (95% CI [93-100]) and 83.2% (95% CI [77.6-87.9]), respectively. Using multiplex real time PCR as a reference, light microscopy and RDT had sensitivity of 58% (95% CI [46.9-68.4] and 67% (95% CI [56.2-76.7]); and 100% (95% CI [98-100] and 98.9 (95% CI (96-99.9), respectively. Substantial level of agreement was reported between microscopy and multiplex real time PCR results with kappa value of 0.65. Conclusions: Multiplex real time PCR had an advanced performance in parasite detection and species identification on febrile patients’ samples than did microscopy and RDT in low malaria transmission settings. It is highly sensitive malaria diagnostic method that can be used in malaria elimination program, particularly for community based epidemiological samples. Although microscopy and RDT had reduced performance when compared to multiplex real time PCR, still had an acceptable performance in diagnosis of malaria cases on patient samples at clinical facilities.

scholarly journals Clinical performance of the point-of-care cobas Liat for detection of SARS-CoV-2 in 20 minutes: A multicenter study

2020 ◽  
Author(s):  
Glen Hansen ◽  
Jamie Marino ◽  
Zi-Xuan Wang ◽  
Kathleen G. Beavis ◽  
John Rodrigo ◽  
...  
Keyword(s):  
Influenza A ◽  
Clinical Performance ◽  
Point Of Care ◽  
Nasopharyngeal Swab ◽  
Molecular Testing ◽  
Rt Pcr ◽  
Percent Agreement ◽  
Individual Care ◽  
Pcr Technology ◽  
Acid Test

Background: Highly accurate testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the point of care (POC) is an unmet diagnostic need in emergency care and time-sensitive outpatient care settings. Reverse transcription-polymerase chain reaction (RT-PCR) technology is the gold-standard for SARS-CoV-2 diagnostics. Methods: We performed a multi-site United States (US) study comparing the clinical performance of the first US Food and Drug Administration (FDA) authorized POC RT-PCR test for detection of SARS-CoV-2 in 20 minutes, the cobas® Liat SARS-CoV-2 & Influenza A/B nucleic acid test, to the most widely used RT-PCR laboratory test, the cobas® 68/8800 SARS-CoV-2 test. Results: Clinical nasopharyngeal swab specimens from 444 patients with 357 evaluable specimens at five US clinical laboratories were enrolled from September 21, 2020 to October 23, 2020. The overall agreement between the Liat and 68/8800 systems for SARS-CoV-2 diagnostics was 98.6% (352/357). Using Liat, positive percent agreement for SARS-CoV-2 was 100% (162/162) and the negative percent agreement was 97.4% (190/195). Conclusion: The Liat is an RT-PCR POC test that provides highly accurate SARS-CoV-2 results in 20 minutes with equivalent performance to high-throughput laboratory molecular testing. Rapid RT-PCR testing at the POC can enable more timely infection control and individual care decisions for Coronavirus Disease 2019.

scholarly journals Clinical evaluation of BD Veritor SARS-CoV-2 point-of-care test performance compared to PCR-based testing and versus the Sofia 2 SARS Antigen point-of-care test.

2020 ◽  
Author(s):  
Stephen Young ◽  
Stephanie Taylor ◽  
Catherine Cammarata ◽  
Celine Roger-Dalbert ◽  
Amanda Montano ◽  
...  
Keyword(s):  
Test Performance ◽  
Rapid Detection ◽  
Clinical Performance ◽  
Point Of Care ◽  
Point Estimate ◽  
Pcr Assay ◽  
Point Of Care Test ◽  
Nasal Swabs ◽  
High Degree ◽  
Antigen Testing

Objectives The clinical performance of the BD Veritor System for Rapid Detection of SARS-CoV-2 antigen (Veritor), a chromatographic immunoassay that detects the SARS-CoV-2 nucleocapsid antigen as a point-of-care test, was evaluated on nasal specimens from individuals with COVID-19 symptoms. Methods and Materials Two studies were completed to determine clinical performance. In the first study, nasal specimens and either nasopharyngeal or oropharyngeal specimens from 251 participants with COVID-19 symptoms (<=7 days from symptom onset [DSO]), >=18 years of age, were utilized to compare Veritor with the Lyra SARS-CoV-2 PCR Assay (Lyra). In the second study, nasal specimens from 361 participants with COVID-19 symptoms (<=5 DSO), >=18 years of age, were utilized to compare performance of Veritor to that of the Sofia 2 SARS Antigen FIA test (Sofia 2). Positive, negative, and overall percent agreement (PPA, NPA, and OPA, respectively) were the primary outcomes. Results In study 1, PPA for Veritor, compared to Lyra, ranged from 81.8%-87.5% for 0-1 through 0-6 DSO ranges. In study 2, Veritor had a PPA, NPA, and OPA of 97.4%, 98.1%, and 98.1%, respectively, with Sofia 2. Discordant analysis showed one Lyra positive missed by Veritor and five Lyra positives missed by Sofia 2; one Veritor positive result was negative by Lyra. Conclusions Veritor met FDA-EUA acceptance criteria for SARS-CoV-2 antigen testing (>=80% PPA point estimate) for the 0-5 and 0-6 DSO ranges. Veritor and Sofia 2 showed a high degree of agreement for SARS-CoV-2 detection. The Veritor test should facilitate rapid and reliable results for COVID-19 diagnosis utilizing easy-to-collect nasal swabs.

scholarly journals Evaluation of a Hydrogel-Based Diagnostic Approach for the Point-of-Care Based Detection of Neisseria gonorrhoeae

Antibiotics ◽  
2018 ◽  
Vol 7 (3) ◽  
pp. 70 ◽  
Author(s):  
Sumudu Perera ◽  
Ali Taheri ◽  
Nurul Khan ◽  
Rajinder Parti ◽  
Stephanie Stefura ◽  
...  
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Real Time ◽  
Sensitivity And Specificity ◽  
Point Of Care ◽  
Pcr Detection ◽  
Diagnostic Approach ◽  
Rt Pcr ◽  
Neisseria Species ◽  
Point Of Care Test

Eleven primer pairs were developed for the identification of Neisseria gonorrhoeae. The sensitivity and specificity of these primers were evaluated by Real Time (RT)-PCR melt curve analyses with DNA from 145 N. gonorrhoeae isolates and 40 other Neisseria or non-Neisseria species. Three primer pairs were further evaluated in a hydrogel-based RT-PCR detection platform, using DNA extracted from 50 N. gonorrhoeae cultures. We observed 100% sensitivity and specificity in the hydrogel assay, confirming its potential as a point-of-care test (POCT) for N. gonorrhoeae diagnosis.

Sign in / Sign up

Export Citation Format

Share Document