Publication date: Sep 04, 2024
Real-time reverse transcription polymerase chain reaction (RT-PCR), a standard method recommended for the diagnosis of coronavirus disease 2019 (COVID-19) requires 2-4 h to get the result. Although antigen test kit (ATK) is used for COVID-19 screening within 15-30 min, the drawback is its limited sensitivity. Hence, a rapid one-step quadruplex real-time RT-PCR assay: termed ƩS COVID-19 targeting ORF1ab, ORF3a, and N genes of SARS-CoV-2; and Avocado sunblotch viroid (ASBVd) as an internal control was developed. Based on strategies including designing high melting temperature primers with short amplicons, applying a fast ramp rate, minimizing hold time, and reducing the range between denaturation and annealing/extension temperatures; the assay could be accomplished within 25 min. The limit of detection of ORF1ab, ORF3a, and N genes were 1. 835, 1. 310, and 1 copy/reaction, respectively. Validation was performed in 205 combined nasopharyngeal and oropharyngeal swabs. The sensitivity, specificity, positive predictive value, and negative predictive value were 92. 8%, 100%, 100%, and 97. 1%, respectively with 96. 7% accuracy. Cohen’s Kappa was 0. 93. The newly developed rapid real-time RT-PCR assay was highly sensitive, specific, and fast, making it suitable for use as an alternative method to support laboratory diagnosis of COVID-19 in outpatient and emergency departments.
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Semantics
Type | Source | Name |
---|---|---|
disease | MESH | COVID 19 |
disease | IDO | assay |
disease | MESH | ramp |
disease | MESH | emergency |
drug | DRUGBANK | Coenzyme M |
disease | IDO | host |
disease | IDO | ribonucleic acid |
disease | MESH | dyspnea |
disease | MESH | sore throat |
disease | MESH | pneumonia |
disease | MESH | death |
disease | IDO | symptom |
drug | DRUGBANK | Ritonavir |
drug | DRUGBANK | Dexamethasone |
drug | DRUGBANK | Baricitinib |
drug | DRUGBANK | Tocilizumab |
disease | IDO | nucleic acid |
disease | MESH | infection |
disease | MESH | influenza |
disease | MESH | parainfluenza |
drug | DRUGBANK | Proline |
disease | IDO | process |
drug | DRUGBANK | Spinosad |
drug | DRUGBANK | Phenylpropanolamine |
disease | MESH | hepatitis |
disease | MESH | Avian Influenza |
drug | DRUGBANK | Water |
drug | DRUGBANK | Timonacic |
drug | DRUGBANK | Edetic Acid |
disease | IDO | facility |
disease | MESH | viral shedding |
drug | DRUGBANK | Ferrous sulfate anhydrous |
disease | MESH | viral load |
disease | MESH | classical swine fever |
drug | DRUGBANK | Methionine |
disease | IDO | replication |
disease | MESH | tics |
drug | DRUGBANK | Coenzyme A |
pathway | REACTOME | Reproduction |