eS COVID-19 is a rapid high throughput and sensitive one-step quadruplex real-time RT-PCR assay.

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.

Open Access PDF

Concepts Keywords
Avocado Adult
Coronavirus COVID-19
Fast COVID-19
Outpatient Female
Pcr High throughput
Humans
Male
Middle Aged
Nasopharynx
Polyproteins
Polyproteins
Rapid
Real time RT-PCR
RNA, Viral
RNA, Viral
SARS-CoV-2
SARS-CoV-2
Sensitivity and Specificity
Viral Proteins
Viral Proteins

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

Original Article

(Visited 1 times, 1 visits today)