Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2.

Publication date: Jul 01, 2025

Early diagnosis of pathogens is key to reducing their spreading and to preventing severe health risks. The COVID-19 pandemic showed us the need for rapid point-of-care tests. Here, we describe the preparation of an amplification master mix for point-of-care diagnostics. Therefore, two off-patent amplification enzymes were designed, expressed and purified. The preparation of the key components enables independence from delivery issues, manufacturer portfolio and product information’s. For long-term storage and cold-free transport, our fabricated amplification mix was lyophilised. Finally, we applied our lyophilised master mix on an integrated point-of-care diagnostic system and could detect 10 copies/μL COVID-19 RNA. The combination of stable, cold-free reagents with the mobile and low-cost device will allow molecular diagnostics of pathogens in a field or home setting.

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Concepts Keywords
Biotechnol COVID-19
Covid Freeze Drying
Independence Humans
Pandemic lyophilisation
Transcriptase Molecular Diagnostic Techniques
Point-of-Care Systems
Point-of-Care Testing
point‐of‐care
pulse controlled amplification
recombinant enzymes
RNA-Directed DNA Polymerase
RNA-Directed DNA Polymerase
RNA, Viral
RNA, Viral
RT‐qPCR
SARS-CoV-2
SARS‐CoV‐2

Semantics

Type Source Name
disease MESH COVID-19 pandemic
disease MESH infectious diseases
disease MESH viral infections
drug DRUGBANK Gold
disease MESH infection
pathway REACTOME Reproduction
drug DRUGBANK Pidolic Acid
disease IDO site
drug DRUGBANK Glycerin
disease IDO protein
drug DRUGBANK Kanamycin
drug DRUGBANK Isopropyl beta-D-thiogalactopyranoside
disease IDO bacteria
drug DRUGBANK Tromethamine
drug DRUGBANK Lysozyme
drug DRUGBANK Imidazole
drug DRUGBANK Iron
drug DRUGBANK Coenzyme M
drug DRUGBANK Heparin
drug DRUGBANK Flunarizine
disease IDO assay
drug DRUGBANK Edetic Acid
drug DRUGBANK Potassium Chloride
drug DRUGBANK Water
drug DRUGBANK Trehalose
drug DRUGBANK Gelatin
drug DRUGBANK Silicon dioxide
drug DRUGBANK Adamantane
drug DRUGBANK Indium
drug DRUGBANK Zinc
drug DRUGBANK Nitrogen
drug DRUGBANK Aluminium
drug DRUGBANK Amino acids
drug DRUGBANK Sodium lauryl sulfate
drug DRUGBANK Creatinolfosfate
drug DRUGBANK Indoleacetic acid
drug DRUGBANK Phosphate ion
disease IDO reagent
disease MESH Mycobacterium ulcerans Infection
disease MESH Neglected Tropical Diseases
disease IDO production
disease IDO deoxyribonucleic acid
disease IDO blood
disease IDO nucleic acid

Original Article

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