NDLI: A new microfluidic approach for the one-step capture, amplification and label-free quantification of bacteria from raw samples

Content Provider	Royal Society of Chemistry (RSC)
Author	Tabnaoui, Sanae Joshi, Lokesh Malaquin, Laurent Deegan, Shane Bilkova, Zuzana Bendali, Amel Alexandre, Lucile Srbova, Jana Descroix, Stéphanie Viovy, Jean-Louis Pereiro, Iago Dupuy, Bruno
Copyright Year	2017
Abstract	A microfluidic method to specifically capture and detect infectious bacteria based on immunorecognition and proliferative power is presented. It involves a microscale fluidized bed in which magnetic and drag forces are balanced to retain antibody-functionalized superparamagnetic beads in a chamber during sample perfusion. Captured cells are then cultivated in situ by infusing nutritionally-rich medium. The system was validated by the direct one-step detection of Salmonella Typhimurium in undiluted unskimmed milk, without pre-treatment. The growth of bacteria induces an expansion of the fluidized bed, mainly due to the volume occupied by the newly formed bacteria. This expansion can be observed with the naked eye, providing simple low-cost detection of only a few bacteria and in a few hours. The time to expansion can also be measured with a low-cost camera, allowing quantitative detection down to 4 cfu (colony forming unit), with a dynamic range of 100 to 107 cfu ml−1 in 2 to 8 hours, depending on the initial concentration. This mode of operation is an equivalent of quantitative PCR, with which it shares a high dynamic range and outstanding sensitivity and specificity, operating at the live cell rather than DNA level. Specificity was demonstrated by controls performed in the presence of a 500× excess of non-pathogenic Lactococcus lactis. The system's versatility was demonstrated by its successful application to the detection and quantitation of Escherichia coli O157:H15 and Enterobacter cloacae. This new technology allows fast, low-cost, portable and automated bacteria detection for various applications in food, environment, security and clinics.
Starting Page	1329
Ending Page	1336
Page Count	8
File Format	HTM / HTML PDF
ISSN	20416520
Volume Number	8
Issue Number	2
Journal	Chemical Science
DOI	10.1039/c6sc03880h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Subscribed
Subject Keyword	Lactococcus lactis Dynamic range Salmonella enterica subsp. enterica Superparamagnetism Colony-forming unit Enterobacter Perfusion DNA Bacteria High-dynamic-range imaging List of observatory codes Escherichia coli O157:H7 Fluidized bed Real-time polymerase chain reaction
Content Type	Text
Resource Type	Article
Subject	Chemistry

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Colonization of Arabidopsis thaliana with Salmonella enterica and enterohemorrhagic Escherichia coli O157:H7 and competition by Enterobacter asburiae.

A lactose fermentation product produced by Lactococcus lactis subsp. lactis, acetate, inhibits the motility of flagellated pathogenic bacteria.

Survival and growth of Escherichia coli O157:H7, Yersinia enterocolitica and Salmonella enteritidis on decontaminated and untreated meat.

Lactoferrin as an antimicrobial against Salmonella enterica and Escherichia coli O157:H7 in raw milk.

PCR amplification of the gene acmA differentiates Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris.

Lactococcus lactis subsp. lactis† (1998)

Lactococcus lactis subsp. Lactis† (1998).

Phenotypic and Genotypic Characterization of Nisin-Producing Lactococcus Lactis Subsp. Lactis YB23 Isolated from Raw Milk in Turkey

Zahter (Thymbra spicata L.) Bitkisi Ekstraktının, Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium ve Campylobacter jejuni Üzerindeki Antibakteriyel Etkileri

A new microfluidic approach for the one-step capture, amplification and label-free quantification of bacteria from raw samples

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Colonization of Arabidopsis thaliana with Salmonella enterica and enterohemorrhagic Escherichia coli O157:H7 and competition by Enterobacter asburiae.

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Survival and growth of Escherichia coli O157:H7, Yersinia enterocolitica and Salmonella enteritidis on decontaminated and untreated meat.

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PCR amplification of the gene acmA differentiates Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris.

Lactococcus lactis subsp. lactis† (1998)

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A new microfluidic approach for the one-step capture, amplification and label-free quantification of bacteria from raw samples