Microdroplet-based multiplex PCR on chip to detect foodborne bacteria producing biogenic amines

被引:17
作者
Sciancalepore, Anna Giovanna [1 ]
Mele, Elisa [1 ]
Arcadio, Valentina [2 ,3 ]
Reddavide, Francesco [2 ,4 ]
Grieco, Francesco [5 ]
Spano, Giuseppe [6 ]
Lucas, Patrick [7 ]
Mita, Giovanni [5 ]
Pisignano, Dario [2 ]
机构
[1] Fdn Ist Italian Tecnol UniLe, Ctr Biomol Nanotechnol, I-73010 Lecce, Italy
[2] Ist Nanosci CNR, NNL, I-73100 Lecce, Italy
[3] Univ Salento, Fac Sci Matemat Fis & Nat, I-73100 Lecce, Italy
[4] Univ Bari, Fac Sci Biotecnol, I-70125 Bari, Italy
[5] CNR Ist Sci Prod Alimentari, I-73100 Lecce, Italy
[6] Dipartimento Sci Agr Alimenti & Ambiente, I-71100 Foggia, Italy
[7] Univ Bordeaux, ISVV, INRA, UMR Enol, F-33882 Villenave Dornon, France
关键词
Biogenic amines; Microfluidics; Multiplex-PCR; Food safety; LACTOBACILLUS-BREVIS; RAPID DETECTION; ACID BACTERIA; WINE; FOODS; IDENTIFICATION; AMPLIFICATION; GENES;
D O I
10.1016/j.fm.2013.02.010
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The development of fast, reliable and culture-independent molecular tools to detect bacteria producing biogenic amines deserves the attention of research and ultimately of the food industry in order to protect consumers' health. Here we present the application of a simple, low-cost, fast and sensitive method to perform microdroplet-based multiplex PCR, directly on a food matrix, for the simultaneous detection of bacterial genes involved in biogenic amine biosynthesis. After inoculating wine with Lactobacillus brevis IOEB 9809, cell lysis and DNA amplification are performed in one single step, without preliminary nucleic acid extraction or purification treatments. The assay is performed in about 30 min, requiring 150 nL of starting sample and it enables the detection of down to 15 bacterial cells. With respect to traditional culture techniques, the speed, the simplicity and the cheapness of this procedure allow an effective monitoring of microbial cells during food-making and processing. (C) 2013 Elsevier Ltd. All rights reserved.
引用
收藏
页码:10 / 14
页数:5
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