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While pooling enhances RT\u2010PCR throughput, reducing costs and resources, it dilutes positive samples with negative ones, lowering sensitivity and increasing false negatives. This study proposes a new method to address the trade\u2010off between pool sizes and RT\u2010PCR accuracy. This method integrates large\u2010scale pooling with sample enrichment using a nano\u2010hybrid membrane, preventing the pooling\u2010induced decrease in viral concentration and maintaining cycle threshold (Ct) values close to individual positive samples. The nano\u2010hybrid membrane, named SIMPLE (streamlined, simple, and inexpensive method for preconcentration, lysis, and nucleic acid extraction), comprises layered red blood cell membranes, polyethersulfone, and silica membranes. Using SIMPLE, a Ct value reduction to\u00a0\u22482.6 is demonstrated in pooled COVID\u201019 samples with a pool size of 6 and found Ct values from larger pool sizes (8, 16, 32, 64, and 128) comparable to individual positive samples.<\/jats:p>","DOI":"10.1002\/advs.202408771","type":"journal-article","created":{"date-parts":[[2025,1,21]],"date-time":"2025-01-21T00:41:41Z","timestamp":1737420101000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Enhancing RT\u2010PCR Throughput and Sensitivity through Large\u2010Scale Sample Pooling Using a Nano\u2010Hybrid Membrane"],"prefix":"10.1002","volume":"12","author":[{"given":"Na Eun","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Electrical Engineering Kwangwoon University  Seoul 01897 Republic of Korea"},{"name":"Department of Biotechnology College of Life Sciences and Biotechnology Korea University  Anam\u2010dong, Seongbuk\u2010gu Seoul 02841 Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kang Hyeon","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Electrical Engineering Kwangwoon University  Seoul 01897 Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ji Hye","family":"Hong","sequence":"additional","affiliation":[{"name":"Department of Electrical Engineering Kwangwoon University  Seoul 01897 Republic of Korea"},{"name":"School of Biomedical Engineering Korea University  Seoul 02841 Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seungmin","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Electrical Engineering Kwangwoon University  Seoul 01897 Republic of Korea"},{"name":"School of Biomedical Engineering Korea University  Seoul 02841 Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeong Soo","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Electrical Engineering Kwangwoon University  Seoul 01897 Republic of Korea"},{"name":"School of Mechanical Engineering Korea University  145 Anam\u2010ro, Seoungbuk\u2010gu Seoul 02841 Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dohwan","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Electrical Engineering Kwangwoon University  Seoul 01897 Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dae Sung","family":"Yoon","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering Korea University  Seoul 02841 Republic of Korea"},{"name":"Interdisciplinary Program in Precision Public Health Korea University  Seoul 02841 Republic of Korea"},{"name":"Astrion Inc.  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