A microfluidic droplet digital PCR for simultaneous detection of pathogenic Escherichia coli O157 and Listeria monocytogenes

Bian, Xiaojun; Fu, Jing; Li, Gang; Fan, Xiaoyun; Jia, Chunping; Zhou, Hongbo; Jin, Qinghui; Zhao, Jianlong

doi:10.1016/j.bios.2015.07.016

Cited by 155 publications

(93 citation statements)

References 47 publications

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“…Given that significant variability has been observed in the removal rates of indicator bacteria by MBRs in previous studies (Zanetti et al 2010), a systematic assessment based on comprehensive molecular-based detection is therefore needed to determine the removal efficacies of aerobic and anaerobic MBRs. In particular, the use of high-detection sensitivity methods such as high-throughput sequencing and digital PCR would be useful in addressing the removal of pathogens by MBRs (Bian et al 2015; Cai and Zhang 2013). …”

Section: Introductionmentioning

confidence: 99%

Molecular-based detection of potentially pathogenic bacteria in membrane bioreactor (MBR) systems treating municipal wastewater: a case study

Harb

Hong

2016

Environ Sci Pollut Res

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Although membrane bioreactor (MBR) systems provide better removal of pathogens compared to conventional activated sludge processes, they do not achieve total log removal. The present study examines two MBR systems treating municipal wastewater, one a full-scale MBR plant and the other a lab-scale anaerobic MBR. Both of these systems were operated using microfiltration (MF) polymeric membranes. High-throughput sequencing and digital PCR quantification were utilized to monitor the log removal values (LRVs) of associated pathogenic species and their abundance in the MBR effluents. Results showed that specific removal rates vary widely regardless of the system employed. Each of the two MBR effluents’ microbial communities contained genera associated with opportunistic pathogens (e.g., Pseudomonas, Acinetobacter) with a wide range of log reduction values (< 2 to >5.5). Digital PCR further confirmed that these bacterial groups included pathogenic species, in several instances at LRVs different than those for their respective genera. These results were used to evaluate the potential risks associated both with the reuse of the MBR effluents for irrigation purposes and with land application of the activated sludge from the full-scale MBR system.Electronic supplementary materialThe online version of this article (doi:10.1007/s11356-016-8211-y) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Molecular-based detection of potentially pathogenic bacteria in membrane bioreactor (MBR) systems treating municipal wastewater: a case study

Harb

Hong

2016

Environ Sci Pollut Res

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“…was the most frequent studied pathogen followed by Listeria spp. Other microorganisms reported were E. coli O157:H7, E. coli, Bacilus spp., Campylobacter spp., Vibrio spp and Shigela spp due to the fact of some of them having the capacity to produce dangerous toxins [16] . The most employed technique, in both simple and multiplex detection methods, was PCR.…”

Section: Discussionmentioning

confidence: 99%

“…e Shigela spp. devido a capacidade de alguns deles produzir perigosas toxina, podendo colocar em risco a saúde da população [16] . A técnica predominantemente empregada tanto em mé-todos de detecção simples quanto em multiplex foi a de PCR.…”

Section: Discussionunclassified

Comparison between Listeria monocytogenes single and multipathogen detection methods

Souza¹,

Ferreira²,

Pasquini-Netto³

et al. 2017

BBR

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143 Biomedical and Biopharmaceutical Research J o r n a l d e I n v e s t i g a ç ã o B i o m é d i c a e B i o f a r m a c ê u t i c a Comparison between Listeria monocytogenes single and multipathogen detection methods Comparação entre os métodos de detecção de Listeria monocytogenes simples e multipatógenos AbstractMultipathogen detection methods have been used to provide a broad range of microorganism detection. We aimed to identify the most searched pathogens in L. monocytogenes detection methods and evaluate if there are sensitivity and specificity differences between single and multipathogen methods. A systematic review was performed by including studies carried out in order to detect L. monocytogenes in a broad range of food sample. A total of 2770 records were retrieved, of which 191 were selected. The majority of the studies (104) presented multipathogen detection, other 87 searched for L. monocytogenes specifically. From the studies for multipathogen detection; other Listeria species (19.4%), Salmonella spp. (21.6%), E. coli O157:H7 (13.2%), Staphylococcus spp. (9.7%), E. coli (5.7%) were more frequently reported. Sensitivity and specificity calculations were derived from only 26 studies, because they compared their data with an official method. Sensitivity and specificity values were close to 100%, showing that others characteristics such as time and cost should be considered to evaluate alternative methods in further research. As a conclusion, evidence generated regarding L. monocytogenes identification methods contribute to method improvements and listeriosis control.Keywords: L. monocytogenes, methods, pathogens, sensitivity, specificity. ResumoOs métodos de detecção para múltiplos patógenos têm sido usados para fornecer uma ampla faixa de detecção de microrganismos. Objetivou-se identificar os patógenos mais buscados nos métodos de detecção para L. monocytogenes e avaliar se existem diferenças nas taxas de sensibilidade e especificidade entre os métodos que buscaram identificar somente L. monocytogenes e os para múltiplos patógenos. Uma revisão sistemática foi realizada incluindo estudos desenvolvidos para detectar L. monocytogenes em amostras alimentares.

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“…Mineral oils and perfluorinated oils (such as HFE-7500, FC-40 and FC-70) are most two frequently used. When using mineral oils droplets can be efficiently stabilised by Span 80 (sorbitan monooleate) and Abil EM 90 (a non-ionic, silicone-based emulsifier) (Williams et al 2006;Schütze et al 2011;Bian et al 2015). Nevertheless, due to the prevalence of fluorinated oils as carrier fluids (because of their excellent biocompatibility and high gas permeabilities), fluorosurfactants (perfluoropolyethers containing Left: Schematics and false-coloured images of devices.…”

Section: Droplet Stabilitymentioning

confidence: 99%

From single-molecule detection to next-generation sequencing: microfluidic droplets for high-throughput nucleic acid analysis

Ding

Choo

deMello

2017

Microfluid Nanofluid

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A microfluidic droplet digital PCR for simultaneous detection of pathogenic Escherichia coli O157 and Listeria monocytogenes

Cited by 155 publications

References 47 publications

Molecular-based detection of potentially pathogenic bacteria in membrane bioreactor (MBR) systems treating municipal wastewater: a case study

Molecular-based detection of potentially pathogenic bacteria in membrane bioreactor (MBR) systems treating municipal wastewater: a case study

Comparison between Listeria monocytogenes single and multipathogen detection methods

From single-molecule detection to next-generation sequencing: microfluidic droplets for high-throughput nucleic acid analysis

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