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dc.contributor.authorTsou, J.-H.
dc.contributor.authorLeng, Q.
dc.contributor.authorJiang, F.
dc.date.accessioned2020-03-23T15:54:16Z
dc.date.available2020-03-23T15:54:16Z
dc.date.issued2020
dc.identifier.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85081213709&doi=10.3390%2fdiagnostics10020114&partnerID=40&md5=7c683e1316041011d4c85c1de400ee08
dc.identifier.urihttp://hdl.handle.net/10713/12319
dc.description.abstractThe detection of EGFR mutations in circulating cell-free DNA can enable personalized therapy for cancer. The current techniques for detecting circulating EGFR mutations are expensive and time-consuming with moderate sensitivity. Emerging CRISPR is revolutionizing medical diagnostics and showing a great promise for nucleic acid detection. This study aims to develop CRISPR-Cas12a as a simple test to sensitively detect circulating EGFR mutations in plasma. Serially diluted samples of DNA containing heterozygous EGFR mutations (L858R and T790M) in wild-type genomic DNA are concurrently tested for the mutations by a CRISPR-Cas12a system and droplet digital PCR (ddPCR). The CRISPR-Cas12a system can detect both L858R and T790M with a limit of detection of 0.005% in less than three hours. ddPCR detects the mutations with a limit of detection of 0.05% for more than five hours. Plasma samples of 28 lung cancer patients and 20 cancer-free individuals are tested for the EGFR mutations by CRISPR-Cas12a system and ddPCR. The CRISPR-Cas12a system could detect L858R in plasma of two lung cancer patients whose tissue biopsies are positive for L858R, and one plasma sample of three lung cancer patients whose tissue biopsies are positive for T790M. ddPCR detects L858R in the same two plasm samples, however, does not detect T790M in any of the plasma samples. This proof of principle study demonstrates that the CRISPR-Cas12a system could rapidly and sensitively detect circulating EGFR mutations, and thus, has potential prognostic or therapeutic implications. Copyright 2020 by the authors.en_US
dc.description.sponsorshipThis work was supported in part by the Geaton and JoAnn DeCesaris Family Foundation and University of Maryland Marlene and Stewart Greenebaum Comprehensive Cancer Center Pilot Grant Program (F.J.).en_US
dc.description.urihttps://doi.org/10.3390/diagnostics10020114en_US
dc.language.isoen_USen_US
dc.publisherMDPI AGen_US
dc.relation.ispartofDiagnostics
dc.subjectCanceren_US
dc.subjectCRISPRen_US
dc.subjectEGFRen_US
dc.subjectLiquid biopsyen_US
dc.subjectMutationsen_US
dc.subjectPlasmaen_US
dc.titleA CRISPR test for rapidly and sensitively detecting circulating EGFR mutationsen_US
dc.typeArticleen_US
dc.identifier.doi10.3390/diagnostics10020114


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