ArticleInternational journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases2021
Analytical sensitivity and clinical performance of a triplex RT-qPCR assay using CDC N1, N2, and RP targets for SARS-CoV-2 diagnosis.
Article in International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers, 1 of them a synthesis that pooled it.
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Who cites it
29 citing papers in PubMed, 1 synthesis or guideline pooled it, 40 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
- Coinfections and superinfections in critically ill COVID-19 patients in Ecuador: a cross-sectional study of bacterial and viral pathogens.Frontiers in public health · 2025Article
- Fast and sensitive CRISPR detection by minimized interference of target amplification.Nature chemical biology · 2024Article
- Descriptive analysis of Cycle Threshold in patients with hematologic malignancies infected with SARS-COV-2.Revista de la Facultad de Ciencias Medicas (Cordoba, Argentina) · 2023Observational
- Coinfection of SARS-CoV-2 with other respiratory pathogens in outpatients from Ecuador.Frontiers in public health · 2023Article
- Fast, cheap and sensitive: Homogenizer-based RNA extraction free method for SARS-CoV-2 detection by RT-qPCR.Frontiers in cellular and infection microbiology · 2023Article
- The quality of commercial SARS-CoV-2 nucleic acid tests in Ecuador: lessons from COVID-19 pandemic for advancing social equity through microbiology.Frontiers in cellular and infection microbiology · 2023Article
- Performance Evaluation of Developed Bangasure™ Multiplex rRT-PCR Assay for SARS-CoV-2 Detection in Bangladesh: A Blinded Observational Study at Two Different Sites.Diagnostics (Basel, Switzerland) · 2022Article
- Improving the Detection Sensitivity of a New Rapid Diagnostic Technology for Severe Acute Respiratory Syndrome Coronavirus 2 Using a Trace Amount of Saliva.Diagnostics (Basel, Switzerland) · 2022Article
- COVID-19 in Children: Correlation Between Epidemiologic, Clinical Characteristics, and RT-qPCR Cycle Threshold Values.The Pediatric infectious disease journal · 2022Observational
- SARS-CoV-2 Viral Load Analysis at Low and High Altitude: A Case Study from Ecuador.International journal of environmental research and public health · 2022Article
- An Overview of SARS-CoV-2 Molecular Diagnostics in Europe.Clinics in laboratory medicine · 2022Review
- Analytical sensitivity and clinical performance of "COVID-19 RT-PCR Real TM FAST (CY5) (ATGen, Uruguay) and "ECUGEN SARS-CoV-2 RT-qPCR" (UDLA-STARNEWCORP, Ecuador)": High quality-low cost local SARS-CoV-2 tests for South America.PLoS neglected tropical diseases · 2022Article
- Sustained COVID-19 community transmission and potential super spreading events at neglected afro-ecuadorian communities assessed by massive RT-qPCR and serological testing of community dwelling population.Frontiers in medicine · 2022Article
- Diagnostic Performance of Seven Commercial COVID-19 Serology Tests Available in South America.Frontiers in cellular and infection microbiology · 2022Article
- BNT162b2 against COVID-19 in Brazil using a test-negative design: Study protocol and statistical analysis plan.PloS one · 2022Article
- Clinical Performance of Three Commercial SARS-CoV-2 Rapid Antigen Tests for Community-Dwelling Individuals in a Tropical Setting.Frontiers in cellular and infection microbiology · 2022Article
- Crucial contribution of the universities to SARS-CoV-2 surveillance in Ecuador: Lessons for developing countries.One health (Amsterdam, Netherlands) · 2021Review
- COVID-19 Community Transmission and Super Spreaders in Rural Villages from Manabi Province in the Coastal Region of Ecuador Assessed by Massive Testing of Community-Dwelling Population.The American journal of tropical medicine and hygiene · 2021Article
- High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization.Scientific reports · 2021Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundSeveral RT-qPCR kits are available for SARS-CoV-2 diagnosis and some have emergency use authorization from the US Food and Drug Administration. In particular, the nCoV19 CDC kit includes two targets for detecting SARS-CoV-2 (N1 and N2) and an RNaseP (RP) target for RNA extraction quality control, all of which are labeled with FAM, and thus three PCR reactions are required per sample.
methodsWe designed a triplex RT-qPCR assay based on nCoV19 primers and probes where N1, N2, and RP are labeled with FAM, HEX, and Cy5, respectively, so only a single PCR reaction is required for each sample for SARS-CoV-2 diagnosis.
resultsIn total, 172 samples were analyzed in both singleplex and triplex assays, where 86 samples tested SARS-CoV-2 negative with both assays, so the triplex assay specificity was 100%. In addition, 86 samples tested SARS-Co-V 2 positive with the singleplex assay and 84 with the triplex assay, so the sensitivity was 97.7%. The limit of detection for the triplex assay was determined as 1000 copies/mL.
conclusionsThis new triplex RT-qPCR assay based on primers and probes from the CDC protocol is highly reliable for SARS-CoV-2 diagnosis, and it could speed up detection and save reagents during the current SARS-CoV-2 testing supplies shortage.
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