Natalie Mutter, Barbara Posch, Yasmin Gillitschka, Nadezhda Kataeva, Birgit Michalek, Daniel Lager, Filip Staniszewski, Alexandra Schilder, Lidiya Osinkina, Maximilian Westenthanner, Joachim Stehr, Federico Buersgens, Johannes R. Peham
{"title":"冻干逆转录酶和聚合酶用于SARS-CoV-2的即时诊断","authors":"Natalie Mutter, Barbara Posch, Yasmin Gillitschka, Nadezhda Kataeva, Birgit Michalek, Daniel Lager, Filip Staniszewski, Alexandra Schilder, Lidiya Osinkina, Maximilian Westenthanner, Joachim Stehr, Federico Buersgens, Johannes R. Peham","doi":"10.1111/1751-7915.70200","DOIUrl":null,"url":null,"abstract":"<p>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.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 7","pages":""},"PeriodicalIF":5.2000,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70200","citationCount":"0","resultStr":"{\"title\":\"Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2\",\"authors\":\"Natalie Mutter, Barbara Posch, Yasmin Gillitschka, Nadezhda Kataeva, Birgit Michalek, Daniel Lager, Filip Staniszewski, Alexandra Schilder, Lidiya Osinkina, Maximilian Westenthanner, Joachim Stehr, Federico Buersgens, Johannes R. Peham\",\"doi\":\"10.1111/1751-7915.70200\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>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.</p>\",\"PeriodicalId\":209,\"journal\":{\"name\":\"Microbial Biotechnology\",\"volume\":\"18 7\",\"pages\":\"\"},\"PeriodicalIF\":5.2000,\"publicationDate\":\"2025-07-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70200\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbial Biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.70200\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.70200","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2
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.
期刊介绍:
Microbial Biotechnology publishes papers of original research reporting significant advances in any aspect of microbial applications, including, but not limited to biotechnologies related to: Green chemistry; Primary metabolites; Food, beverages and supplements; Secondary metabolites and natural products; Pharmaceuticals; Diagnostics; Agriculture; Bioenergy; Biomining, including oil recovery and processing; Bioremediation; Biopolymers, biomaterials; Bionanotechnology; Biosurfactants and bioemulsifiers; Compatible solutes and bioprotectants; Biosensors, monitoring systems, quantitative microbial risk assessment; Technology development; Protein engineering; Functional genomics; Metabolic engineering; Metabolic design; Systems analysis, modelling; Process engineering; Biologically-based analytical methods; Microbially-based strategies in public health; Microbially-based strategies to influence global processes