کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5031380 1470933 2018 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A nanoparticle-based method for culture-free bacterial DNA enrichment from whole blood
ترجمه فارسی عنوان
یک روش مبتنی بر نانوذرات برای غنی سازی DNA باکتریایی بدون کریستالی از کل خون
کلمات کلیدی
POCT؛ تشخیص باکتری سپسیس؛ خون؛ نانوذرات مغناطیسی؛ qPCR
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی

Point-of-care (POC) diagnostics are one of the quick and sensitive detection approaches used in current clinical applications, but always face a performance tradeoff between time-to-result and assay sensitivity. One critical setting where these limitations are evident is the detection of sepsis, where 6-10 mL of whole blood may contain as little as one bacterial colony forming unit (cfu). The large sample volume, complex nature of the sample and low analyte concentration necessitates signal enhancement using culture-based or molecular amplification techniques. In the time-critical diagnosis of sepsis, waiting for up to 24 h to produce sufficient DNA for analysis is not possible. As a consequence, there is a need for integrated sample preparation methods that could enable shorter detection times, whilst maintaining high analytical performance. We report the development of a culture-free bacterial enrichment method to concentrate bacteria from whole blood in less than 3 h. The method relies on triple-enrichment steps to magnetically concentrate bacterial cells and their DNA with a 500-fold reduction in sample volume (from 10 to 0.02 mL). Using this sample preparation method, sensitive qPCR detection of the extracted S. aureus bacterial DNA was achieved with a detection limit of 5±0.58 cfu/mL within a total elapsed time of 4 h; much faster than conventional culture-based approaches. The method could be fully automated for integration into clinical practice for point-of-care or molecular detection of bacterial DNA from whole blood.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 99, 15 January 2018, Pages 150-155
نویسندگان
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