کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
7609878 1493403 2017 21 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Sequence-specific DNA solid-phase extraction in an on-chip monolith: Towards detection of antibiotic resistance genes
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Sequence-specific DNA solid-phase extraction in an on-chip monolith: Towards detection of antibiotic resistance genes
چکیده انگلیسی
Antibiotic resistance of bacteria is a growing problem and presents a challenge for prompt treatment in patients with sepsis. Currently used methods rely on culturing or amplification; however, these steps are either time consuming or suffer from interference issues. A microfluidic device was made from black polypropylene, with a monolithic column modified with a capture oligonucleotide for sequence selective solid-phase extraction of a complementary target from a lysate sample. Porous properties of the monolith allow flow and hybridization of a target complementary to the probe immobilized on the column surface. Good flow-through properties enable extraction of a 100 μL sample and elution of target DNA in 12 min total time. Using a fluorescently labeled target oligonucleotide related to Verona Integron-Mediated Metallo-β-lactamase it was possible to extract and detect a 1 pM sample with 83% recovery. Temperature-mediated elution by heating above the duplex melting point provides a clean extract without any agents that interfere with base pairing, allowing various labeling methods or further downstream processing of the eluent. Further integration of this extraction module with a system for isolation and lysis of bacteria from blood, as well as combining with single-molecule detection should allow rapid determination of antibiotic resistance.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1523, 10 November 2017, Pages 309-315
نویسندگان
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