ArticleJournal of clinical virology : the official publication of the Pan American Society for Clinical Virology2020
Multiplexing primer/probe sets for detection of SARS-CoV-2 by qRT-PCR.
Article in Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers, 1 of them a synthesis that pooled it.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
20 citing papers in PubMed, 1 synthesis or guideline pooled it, 39 citations in OpenAlex.
- Diagnostic performances of common nucleic acid tests for SARS-CoV-2 in hospitals and clinics: a systematic review and meta-analysis.The Lancet. Microbe · 2021Pooled it
- Article
- Performance Comparison of a Duplex Implementation of the CDC EUA 2019-nCoV Assay with the Seegene Allplex-SARS-CoV-2 Assay for the Detection of SARS-CoV-2 in Nasopharyngeal Swab Samples.Methods and protocols · 2022Article
- Article
- Strategies for Scaling up SARS-CoV-2 Molecular Testing Capacity.Clinics in laboratory medicine · 2022Review
- Toward a next-generation diagnostic tool: A review on emerging isothermal nucleic acid amplification techniques for the detection of SARS-CoV-2 and other infectious viruses.Analytica chimica acta · 2022Review
- SARS-CoV-2 detection in wastewater using multiplex quantitative PCR.The Science of the total environment · 2021Article
- Review on oxidative stress relation on COVID-19: Biomolecular and bioanalytical approach.International journal of biological macromolecules · 2021Review
- Surveillance of SARS-CoV-2 lineage B.1.1.7 in Slovakia using a novel, multiplexed RT-qPCR assay.Scientific reports · 2021Article
- Specific allelic discrimination of N501Y and other SARS-CoV-2 mutations by ddPCR detects B.1.1.7 lineage in Washington State.Journal of medical virology · 2021Article
- Coronavirus Disease 2019 (COVID-19) Diagnostic Tools: A Focus on Detection Technologies and Limitations.Current issues in molecular biology · 2021Review
- Development and performance evaluation of the first in-house multiplex rRT-PCR assay in Bangladesh for highly sensitive detection of SARS-CoV-2.Journal of virological methods · 2021Article
- Enhanced throughput of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) real-time RT-PCR panel by assay multiplexing and specimen pooling.Journal of virological methods · 2021Article
- Sample pooling strategies for SARS-CoV-2 detection.Journal of virological methods · 2021Review
- Comparative effects of viral-transport-medium heat inactivation upon downstream SARS-CoV-2 detection in patient samples.Journal of medical microbiology · 2021Article
- COVID-19: A challenge for electrochemical biosensors.Trends in analytical chemistry : TRAC · 2021Review
- Assessment of SARS-CoV-2 infectivity of upper respiratory specimens from COVID-19 patients by virus isolation using VeroE6/TMPRSS2 cells.BMJ open respiratory research · 2021Article
- Modifying laboratory testing via home brew during the COVID-19 pandemic.Journal of clinical and translational science · 2021Article
- Analytical sensitivity and clinical performance of a triplex RT-qPCR assay using CDC N1, N2, and RP targets for SARS-CoV-2 diagnosis.International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases · 2021Article
- The estimation of diagnostic accuracy of tests for COVID-19: A scoping review.The Journal of infection · 2020Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe novel respiratory virus SARS-CoV-2, responsible for over 380,000 COVID-19 related deaths, has caused significant strain on healthcare infrastructure and clinical laboratories globally. The pandemic's initial challenges include broad diagnostic testing, consistent reagent supply lines, and access to laboratory instruments and equipment. In early 2020, primer/probe sets distributed by the CDC utilized the same fluorophore for molecular detection - requiring multiple assays to be run in parallel - consuming valuable and limited resources.
methodsNasopharyngeal swabs submitted to UW Virology for SARS-CoV-2 clinical testing were extracted, amplified by our laboratory developed test (LDT) - a CDC-based quantitative reverse transcriptase PCR reaction - and analyzed for agreement between the multiplexed assay. Laboratory- confirmed respiratory infection samples were included to evaluate assay cross-reaction specificity.
resultsTriplexing correctly identified SARS-CoV-2 in 98.4% of confirmed positive or inconclusive patient samples by single-plex LDT (n = 183/186). All 170 SARS-CoV-2 negative samples tested by single-plex LDT were negative by triplexing. Other laboratory-confirmed respiratory infections did not amplify for SARS-CoV-2 in the triplex reaction.
conclusionsMultiplexing two virus-specific gene targets and an extraction control was found to be comparable to running parallel assays independently, while significantly improving assay throughput.
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