tetano
Editor, Senior Moderator
Microb Biotechnol
. 2025 Jul;18(7):e70200.
doi: 10.1111/1751-7915.70200. Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2
Natalie Mutter[SUP] 1 [/SUP], Barbara Posch[SUP] 1 [/SUP], Yasmin Gillitschka[SUP] 1 [/SUP], Nadezhda Kataeva[SUP] 1 [/SUP], Birgit Michalek[SUP] 1 [/SUP], Daniel Lager[SUP] 2 [/SUP], Filip Staniszewski[SUP] 1 [/SUP], Alexandra Schilder[SUP] 1 [/SUP], Lidiya Osinkina[SUP] 3 [/SUP], Maximilian Westenthanner[SUP] 3 [/SUP], Joachim Stehr[SUP] 3 [/SUP], Federico Buersgens[SUP] 3 [/SUP], Johannes R Peham[SUP] 1 [/SUP]
Affiliations
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.
Keywords: RT‐qPCR; SARS‐CoV‐2; lyophilisation; point‐of‐care; pulse controlled amplification; recombinant enzymes.
. 2025 Jul;18(7):e70200.
doi: 10.1111/1751-7915.70200. Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2
Natalie Mutter[SUP] 1 [/SUP], Barbara Posch[SUP] 1 [/SUP], Yasmin Gillitschka[SUP] 1 [/SUP], Nadezhda Kataeva[SUP] 1 [/SUP], Birgit Michalek[SUP] 1 [/SUP], Daniel Lager[SUP] 2 [/SUP], Filip Staniszewski[SUP] 1 [/SUP], Alexandra Schilder[SUP] 1 [/SUP], Lidiya Osinkina[SUP] 3 [/SUP], Maximilian Westenthanner[SUP] 3 [/SUP], Joachim Stehr[SUP] 3 [/SUP], Federico Buersgens[SUP] 3 [/SUP], Johannes R Peham[SUP] 1 [/SUP]
Affiliations
- PMID: 40705463
- DOI: 10.1111/1751-7915.70200
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.
Keywords: RT‐qPCR; SARS‐CoV‐2; lyophilisation; point‐of‐care; pulse controlled amplification; recombinant enzymes.