کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6491364 | 43410 | 2014 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Rapid quantification of Escherichia coli in food and media using bacteriophage T7 amplification and liquid chromatography-multiple reaction monitoring tandem mass spectrometry
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کلمات کلیدی
CFUMRMLC–MS/MS - LC-MS / MSMOI - MEBacteriophage - باکتریوفاژFood biotechnology - بیوتکنولوژی غذاییRapid diagnostics - تشخیص سریعluria-bertani broth - سوپ لووریا برتانیLod - لودlimit of detection - محدودیت تشخیصmultiple reaction monitoring - نظارت چندگانه چندگانهcolony forming units - واحدهای تشکیل دهنده کلنیplaque forming units - واحدهای تشکیل پلاکpfu - پفوmultiplicity of infection - چندین عفونتoptical density - چگالی نوریliquid chromatography tandem mass spectrometry - کروماتوگرافی مایع اسپکترومتری دو طرفه
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Conventional microbiological assays have been a valuable tool for specific enumeration of indicative bacteria of relevance to food and public health, but these culture-based methods are time-consuming and require tedious biochemical and morphological identification. In this work, we exploit the ability of bacteriophage T7 to specifically infect Escherichia coli and amplify nearly a 100-fold in 1-2Â h. Bacteriophage amplification is integrated with liquid chromatography-multiple reaction monitoring tandem mass spectrometry (LC-MRM-MS/MS) for quantitation of phage-specific peptides. Heavy isotopic 15N labeled T7 is introduced as the inoculum phage and internal standard. Quantification is performed by determining the ratio of phage-specific peptides over the internal standard which value is proportional to E. coli numbers. A broad dynamic range of 6-log orders ranging from 3.0Â ÃÂ 103 to 3.0Â ÃÂ 109Â CFU/ml is attained in LB, while between 4.1Â ÃÂ 104-2.7Â ÃÂ 109Â CFU/ml and 1.9Â ÃÂ 103-3.0Â ÃÂ 107Â CFU/ml was enumerated respectively in coconut water and apple juice. With this method, viable E. coli are quantified in 4Â h with a detection limit of 3.0Â ÃÂ 103Â CFU/ml, 4.1Â ÃÂ 104Â CFU/ml and 1.9Â ÃÂ 103Â CFU/ml in LB, coconut water and apple juice, respectively. This method has potential as a rapid tool for detection of fecal contamination during food bioprocessing and distribution to safeguard public health.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 192, Part A, 20 December 2014, Pages 50-58
Journal: Journal of Biotechnology - Volume 192, Part A, 20 December 2014, Pages 50-58
نویسندگان
Mazlina Banu, Daniel Ng, Lu Zheng, Lin-Tang Goh, Xuezhi Bi, Dave Siak-Wei Ow,