Yi, Joon-Yeop Choi, Hyomin Kim, Minyoung Jeong, Yujin Hahn, Ji-Sook Son, Boram Park, Hee Ho Sung, Changmin 2025-07-30T01:00:35Z 2025-07-30T01:00:35Z 2025-07-28 2025-07 https://pubs.kist.re.kr/handle/201004/152879 Advancements in gene and cell therapies introduce "gene and cell doping," requiring efficient and sensitive detection methods. Here, we report a high-throughput multiplexed gene and cell doping analysis (HiMDA) using CRISPR-Cas12a system integrated with blood direct polymerase chain reaction (PCR). Blood direct PCR enables simultaneous amplification of multiple exogenous genes directly from whole-blood samples. Coupled with sequence-specific DNA recognition and fluorescence reporter system, HiMDA achieves multiplexed, on-target detection of doping genes and cells. Our results demonstrate HiMDA's feasibility with only 5 microliters of blood required for the entire 90-minute process. HiMDA exhibits exceptional sensitivity, detecting as few as 2.5 copies of doping target genes from blood-four times more sensitive than current anti-doping standards-and identifying in vivo doping up to 10 days. These findings highlight HiMDA's robust high-throughput, multiplexed capabilities, satisfying the sensitivity and selectivity demands of anti-doping research. HiMDA offers a flexible solution to meet future doping detection challenges. English American Association for the Advancement of Science High-throughput multiplexed gene and cell doping analysis through CRISPR-Cas12a system integrated with blood direct PCR Article 10.1126/sciadv.adv7234 1 Science Advances, v.11, no.28 Science Advances 11 28 Y scie scopus 001525278400011 2-s2.0-105010975145 Multidisciplinary Sciences Science & Technology - Other Topics Article THERAPY ERYTHROPOIETIN