Rapid Concentration and Molecular Enrichment Approach for Sensitive Detection of Escherichia coli and Shigella Species in Potable Water Samples

Maheux, Andrée F.; Bissonnette, Luc; Boissinot, Maurice; Bernier, Jacques; Huppé, Vicky; Picard, François J.; Bérubé, Ève; Bergeron, Michel G.

doi:10.1128/aem.02337-10

Cited by 52 publications

(51 citation statements)

References 28 publications

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“…The uidA gene (coding for ␤-D-glucuronidase) was chosen as the most frequently used target gene for E. coli (24,38,39,40,41). Its specificity, however, has been questioned (27), and therefore, two other promising gene targets, tuf (coding for a protein elongation factor) (25), present in one to two copies, and rodA (coding for a unspecified protein, affecting cell shape and amdinocillin) (26), were also selected. rodA was included as a single-copy gene target and therefore had the potential to act as a molecular proxy for an E. coli cell count.…”

Section: Test Panel Specificity and (Rt-)q-pcr Assaysmentioning

confidence: 99%

“…In order to select a Q-PCR assay and subsequently design RT-Q-PCR assays for E. coli with the highest levels of specificity, sensitivity, and efficiency, a literature review identified three frequently used functional genes for E. coli, uidA (24), tuf (25), and rodA (26) for comparison. The uidA gene (coding for ␤-D-glucuronidase) was chosen as the most frequently used target gene for E. coli (24,38,39,40,41).…”

Section: Test Panel Specificity and (Rt-)q-pcr Assaysmentioning

confidence: 99%

“…Water quality failures due to the survival or regrowth of E. coli within a WDS bring into question the suitability of E. coli as an indicator organism of fecal contamination. To achieve this, we first evaluated a number of current gene targets (24,25,26,27) and Q-PCR assays for E. coli. Furthermore, we expand their use by developing corresponding reverse transcription-quantitative PCR (RT-Q-PCR) assays to detect mRNA and explore their use as indicators of viable organisms.…”

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confidence: 99%

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Survival, Biofilm Formation, and Growth Potential of Environmental and Enteric Escherichia coli Strains in Drinking Water Microcosms

Abberton

Bereschenko

Wielen

et al. 2016

Appl Environ Microbiol

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Escherichia coli is the most commonly used indicator for fecal contamination in drinking water distribution systems (WDS). The assumption is that E. coli bacteria are of enteric origin and cannot persist for long outside their host and therefore act as indicators of recent contamination events. This study investigates the fate of E. coli in drinking water, specifically addressing survival, biofilm formation under shear stress, and regrowth in a series of laboratory-controlled experiments. We show the extended persistence of three E. coli strains (two enteric isolates and one soil isolate) in sterile and nonsterile drinking water microcosms at 8 and 17°C, with T 90 (time taken for a reduction in cell number of 1 log 10 unit) values ranging from 17.4 ؎ 1.8 to 149 ؎ 67.7 days, using standard plate counts and a series of (reverse transcription-)quantitative PCR [(RT-)Q-PCR] assays targeting 16S rRNA, tuf, uidA, and rodA genes and transcripts. Furthermore, each strain was capable of attaching to a surface and replicating to form biofilm in the presence of nutrients under a range of shear stress values (0.6, 2.0, and 4.4 dynes [dyn] cm ؊2 ; BioFlux system; Fluxion); however, cell numbers did not increase when drinking water flowed over the biofilm (P > 0.05 by t test). Finally, E. coli regrowth within drinking water microcosms containing polyethylene PE-100 pipe wall material was not observed in the biofilm or water phase using a combination of culturing and Q-PCR methods for E. coli. The results of this work highlight that when E. coli enters drinking water it has the potential to survive and attach to surfaces but that regrowth within drinking water or biofilm is unlikely. IMPORTANCEThe provision of clean, safe drinking water is fundamental to society. WDS deliver water to consumers via a vast network of pipes. E. coli is used as an indicator organism for recent contamination events based on the premise that it cannot survive for long outside its host. A key public health concern therefore arises around the fate of E. coli on entering a WDS; its survival, ability to form a biofilm, and potential for regrowth. In particular, if E. coli bacteria have the ability to incorporate and regrow within the pipe wall biofilm of a WDS, they could reinoculate the water at a later stage. This study sheds light on the fate of environmental and enteric strains of E. coli in drinking water showing extended survival, the potential for biofilm formation under shear stress, and importantly, that regrowth in the presence of an indigenous microbial community is unlikely. S afe, clean drinking water is the foundation of society. However, even in developed countries, drinking water quality failures occur and have considerable public health impact. In particular, microbiological quality failures can be a significant threat to the supply of drinking water. The exact cause of these water quality malfunctions can be difficult to determine. In public water supplies, inefficient water treatment of the source could result in unwanted micr...

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Section: Test Panel Specificity and (Rt-)q-pcr Assaysmentioning

confidence: 99%

Section: Test Panel Specificity and (Rt-)q-pcr Assaysmentioning

confidence: 99%

mentioning

confidence: 99%

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Survival, Biofilm Formation, and Growth Potential of Environmental and Enteric Escherichia coli Strains in Drinking Water Microcosms

Abberton

Bereschenko

Wielen

et al. 2016

Appl Environ Microbiol

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“…Quality control for each batch of mCP agar was performed, and filter, buffer, and rinse water blanks were included as sterility controls. For performing CRENAME on concentrated water samples, GN-6 filters were aseptically transferred to 3 individual 15-ml polypropylene tubes (Sarstedt, Newton, NC, USA) and processed as described by Maheux et al (27,28).…”

Section: Determination Of the Analytical Limit Of Detection (Lod) Of mentioning

confidence: 99%

“…Colonies presenting discordant results between the phenotype on mCP agar and cpa rtPCR were identified by sequencing the 16S rRNA and cpa genes. Finally, we compared mCP agar and a CRENAME (concentration and recovery of microbial particles, extraction of nucleic acids, and molecular enrichment) procedure plus cpa rtPCR (CRENAME ϩ cpa rtPCR) for their abilities to detect C. perfringens spores in drinking water (27,28).…”

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confidence: 99%

Abilities of the mCP Agar Method and CRENAME Alpha Toxin-Specific Real-Time PCR Assay To Detect Clostridium perfringens Spores in Drinking Water

Maheux

Bérubé

Boudreau

et al. 2013

Appl Environ Microbiol

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We first determined the analytical specificity and ubiquity (i.e., the ability to detect all or most strains) of a Clostridium perfringens-specific real-time PCR (rtPCR) assay based on the cpa gene (cpa rtPCR) by using a bacterial strain panel composed of C. perfringens and non-C. perfringens Clostridium strains. All non-C. perfringens Clostridium strains tested negative, whereas all C. perfringens strains tested positive with the cpa rtPCR, for an analytical specificity and ubiquity of 100%. The cpa rtPCR assay was then used to confirm the identity of 116 putative C. perfringens isolates recovered after filtration of water samples and culture on mCP agar. Colonies presenting discordant results between the phenotype on mCP agar and cpa rtPCR were identified by sequencing the 16S rRNA and cpa genes. Four mCP ؊ /rtPCR ؉ colonies were identified as C. perfringens, whereas 3 mCP ؉ /rtPCR ؊ colonies were identified as non-C. perfringens. The cpa rtPCR was negative with all 51 non-C. perfringens strains and positive with 64 of 65 C. perfringens strains. Finally, we compared mCP agar and a CRENAME (concentration and recovery of microbial particles, extraction of nucleic acids, and molecular enrichment) procedure plus cpa rtPCR (CRENAME ؉ cpa rtPCR) for their abilities to detect C. perfringens spores in drinking water. CRENAME ؉ cpa rtPCR detected as few as one C. perfringens CFU per 100 ml of drinking water sample in less than 5 h, whereas mCP agar took at least 25 h to deliver results. CRENAME ؉ cpa rtPCR also allows the simultaneous and sensitive detection of Escherichia coli and C. perfringens from the same potable water sample. In itself, it could be used to assess the public health risk posed by drinking water potentially contaminated with pathogens more resistant to disinfection.T he presence of Escherichia coli in drinking water indicates a recent fecal contamination, as well as a risk of waterborne gastrointestinal diseases (1). Under some circumstances, however, it has been shown that this microorganism inadequately indicates the presence of viruses and protozoan parasites of human health significance (2). For example, while chlorine in water rapidly inactivates E. coli, it leaves most disinfection-resistant pathogens almost unaffected for several hours (3). According to Payment and Franco (3), Clostridium perfringens is a suitable indicator of human enteric viruses, Giardia cysts, and Cryptosporidium oocysts in finished water and can also be used in the assessment of water treatment processes due to the resistance of Clostridium spores to chlorine. Furthermore, the presence of C. perfringens in water is also associated with fecal contamination, and it has been evaluated and utilized as an alternative indicator of fecal pollution (4-14). Thus, a simple and reliable culture-based method to isolate and enumerate C. perfringens, the membrane filtration method on mCP agar, has been elaborated and evaluated to monitor the presence of C. perfringens in water (15)(16)(17)(18)(19)(20). In the European Union, the mCP a...

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Current Trends on Surface Acoustic Wave Biosensors

Zida

Lin

Khung

2021

Adv Materials Technologies

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The development of rapid biosensing platforms is an integral part for the detection of many diseases, as well as important clinical biomarkers. So far, many new biosensing technologies have been described in literature but many of these novel platforms remain noncommercializable due to challenges in fabrication process, operational conditions as well as other cost concerns. Hence, there is a need to develop efficient and cost‐effective biosensing platforms and of the types available from literature, surface acoustic wave (SAW) biosensors have certain standout attributes. The simple manufacturing requirements of SAW‐based biosensor and its wide operating frequency range have produced promising outcome for bio‐detection and this has rendered this technological platform as a serious contender for point‐of‐care biosensor design. So far, SAW devices are reported for the detection of various bioactive targets of clinical importance and several designs are already commercialized. In this review, the latest development of SAW technologies in recent years for the detection of a broad range of biomolecules is presented and some of the challenges as well as potential future developments of SAW biosensors are highlighted.

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Rapid Concentration and Molecular Enrichment Approach for Sensitive Detection of Escherichia coli and Shigella Species in Potable Water Samples

Cited by 52 publications

References 28 publications

Survival, Biofilm Formation, and Growth Potential of Environmental and Enteric Escherichia coli Strains in Drinking Water Microcosms

Survival, Biofilm Formation, and Growth Potential of Environmental and Enteric Escherichia coli Strains in Drinking Water Microcosms

Abilities of the mCP Agar Method and CRENAME Alpha Toxin-Specific Real-Time PCR Assay To Detect Clostridium perfringens Spores in Drinking Water

Current Trends on Surface Acoustic Wave Biosensors

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