دورية أكاديمية
Multichannel oscillatory-flow multiplex PCR microfluidics for high-throughput and fast detection of foodborne bacterial pathogens.
| العنوان: | Multichannel oscillatory-flow multiplex PCR microfluidics for high-throughput and fast detection of foodborne bacterial pathogens. |
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| المؤلفون: | Zhang C; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China. zhangcs@scnu.edu.cn, Wang H, Xing D |
| المصدر: | Biomedical microdevices [Biomed Microdevices] 2011 Oct; Vol. 13 (5), pp. 885-97. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer US Country of Publication: United States NLM ID: 100887374 Publication Model: Print Cited Medium: Internet ISSN: 1572-8781 (Electronic) Linking ISSN: 13872176 NLM ISO Abbreviation: Biomed Microdevices Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: New York : Springer US Original Publication: Boston : Kluwer Academic Publishers, 1998- |
| مواضيع طبية MeSH: | Food Contamination/*analysis , Foodborne Diseases/*microbiology , High-Throughput Screening Assays/*methods , Microfluidics/*instrumentation , Multiplex Polymerase Chain Reaction/*methods, Animals ; DNA, Bacterial/analysis ; Equipment Design ; Escherichia coli O157/genetics ; Escherichia coli O157/isolation & purification ; Listeria monocytogenes/genetics ; Listeria monocytogenes/isolation & purification ; Meat Products/microbiology ; Microfluidics/methods ; Milk/microbiology ; Musa/microbiology ; Salmonella enterica/genetics ; Salmonella enterica/isolation & purification |
| مستخلص: | In the field of continuous-flow PCR, the amplification throughput in a single reaction solution is low and the single-plex PCR is often used. In this work, we reported a flow-based multiplex PCR microfluidic system capable of performing high-throughput and fast DNA amplification for detection of foodborne bacterial pathogens. As a demonstration, the mixture of DNA targets associated with three different foodborne pathogens was included in a single PCR solution. Then, the solution flowed through microchannels incorporated onto three temperature zones in an oscillatory manner. The effect factors of this oscillatory-flow multiplex PCR thermocycling have been demonstrated, including effects of polymerase concentration, cycling times, number of cycles, and DNA template concentration. The experimental results have shown that the oscillatory-flow multiplex PCR, with a volume of only 5 μl, could be completed in about 13 min after 35 cycles (25 cycles) at 100 μl/min (70 μl/min), which is about one-sixth of the time required on the conventional machine (70 min). By using the presently designed DNA sample model, the minimum target concentration that could be detected at 30 μl/min was 9.8 × 10(-2) ng/μl (278-bp, S. enterica), 11.2 × 10(-2) ng/μl (168-bp, E. coli O157: H7), and 2.88 × 10(-2) ng/μl (106-bp, L. monocytogenes), which corresponds to approximately 3.72 × 10(4) copies/μl, 3.58 × 10(4) copies/μl, and 1.79 × 10(4) copies/μl, respectively. This level of speed and sensitivity is comparable to that achievable in most other continuous-flow PCR systems. In addition, the four individual channels were used to achieve multi-target PCR analysis of three different DNA samples from different food sources in parallel, thereby achieving another level of multiplexing. |
| المشرفين على المادة: | 0 (DNA, Bacterial) |
| تواريخ الأحداث: | Date Created: 20110622 Date Completed: 20120229 Latest Revision: 20110905 |
| رمز التحديث: | 20240513 |
| DOI: | 10.1007/s10544-011-9558-y |
| PMID: | 21691814 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1572-8781 |
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| DOI: | 10.1007/s10544-011-9558-y |