Identification of Viable <i>Helicobacter pylori</i> in Drinking Water Supplies by Cultural and Molecular Techniques

Santiago, Paula Beatriz; Moreno, Yolanda; Ferrús, María Antonia

doi:10.1111/hel.12205

Cited by 73 publications

(91 citation statements)

References 32 publications

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“…E. coli O157:H7 is one of the most common pathogenic bacteria in water with a low infective dose, several researchers (Al‐Qadiri et al, ; Kerr et al, ) found that it was able to survive for long storage time under cold temperatures in bottled water without essential nutrients. Moreover, in many E. coli waterborne outbreaks, the detected numbers were less than 10 4 CFU/100 ml (Banihashemi, Van Dyke, & Huck, ; Li, Xin, & Li, ; Santiago, Moreno, & Ferrus, ). Therefore, the detection of E. coli O157:H7 at low concentration was quite necessary.…”

Section: Discussionmentioning

confidence: 99%

Molecular monitoring of disinfection efficacy of E. coli O157:H7 in bottled purified drinking water by quantitative PCR with a novel dye

Cao

Zhou²,

et al. 2019

J Food Process Preserv

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A new method was developed using a novel dye thiazole orange monoazide (TOMA) combined with quantitative real‐time PCR (qPCR) to detect viable Escherichia coli O157:H7 cells. Different from the commonly used PMA‐qPCR assay that is based on membrane integrity, this TOMA‐qPCR method is based on the concept of metabolic activity. TOMA consists of three components: a nucleic acid‐intercalating moiety, a crosslinkable moiety and a linker. TOMA concentration at 50 μg/ml, 20 min incubation time, and 30 min light exposure time were suggested to use for detecting viable cells. When the inoculum concentration was 103 CFU/100 ml, TOMA‐qPCR assay could completely exclude the effect of dead cells treated with heat, chlorine, or UV. Moreover, TOMA‐qPCR could also be used to detect viable but nonculturable state (VBNC) cells. The result shows that TOMA‐qPCR would be an alternative choice for the detection of viable cells. Practical applications To specifically detect only viable cells is of great importance in most detection of microbial diagnostics, especially for detection of foodborne pathogens such as E. coli O157:H7, which could cause health risk at low concentrations. The existing detection methods have some defects in detecting only viable cells. In this article, a novel dye was developed and combined with qPCR, to monitor sanitizing efficacy of different disinfection methods on E. coli O157:H7 at low concentrations. When detecting viable E. coli in bottled water, the established TOMA‐qPCR assay completely exclude the effect of dead cells when the inoculated concentration is 103 CFU/100 ml. This method is also suitable for the detection of VBNC samples. Therefore, this method can be used to detect pathogenic bacteria with low contamination under extreme conditions such as low temperature, oligotrophic, strong radiation, and so on.

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Section: Discussionmentioning

confidence: 99%

Molecular monitoring of disinfection efficacy of E. coli O157:H7 in bottled purified drinking water by quantitative PCR with a novel dye

Cao

Zhou²,

et al. 2019

J Food Process Preserv

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show abstract

“…PMA-PCR was first reported to effectively exclude the signal of dead bacteria (Nocker et al, 2006; Josefsen et al, 2010). Later, this approach was adapted by many scientists to detect viable cells of various foodborne pathogens (Cawthorn and Witthuhn, 2008; Josefsen et al, 2010; Liang et al, 2011; van Frankenhuyzen et al, 2011; Banihashemi et al, 2012; Li and Chen, 2012, 2013; Mamlouk et al, 2012; Soejima et al, 2012; Dinu and Bach, 2013; Singh et al, 2013; Yang Y. et al, 2013; Ditommaso et al, 2015; Li et al, 2015; Santiago et al, 2015; Cattani et al, 2016). The majority of the studies demonstrated that PMA-PCR effectively suppressed the signal of DNA from the dead cells.…”

Section: Limitations Of the Pma Approach And The Remedies To Address mentioning

confidence: 99%

Advances and Challenges in Viability Detection of Foodborne Pathogens

et al. 2016

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Foodborne outbreaks are a serious public health and food safety concern worldwide. There is a great demand for rapid, sensitive, specific, and accurate methods to detect microbial pathogens in foods. Conventional methods based on cultivation of pathogens have been the gold standard protocols; however, they take up to a week to complete. Molecular assays such as polymerase chain reaction (PCR), sequencing, microarray technologies have been widely used in detection of foodborne pathogens. Among molecular assays, PCR technology [conventional and real-time PCR (qPCR)] is most commonly used in the foodborne pathogen detection because of its high sensitivity and specificity. However, a major drawback of PCR is its inability to differentiate the DNA from dead and viable cells, and this is a critical factor for the food industry, regulatory agencies and the consumer. To remedy this shortcoming, researchers have used biological dyes such as ethidium monoazide and propidium monoazide (PMA) to pretreat samples before DNA extraction to intercalate the DNA of dead cells in food samples, and then proceed with regular DNA preparation and qPCR. By combining PMA treatment with qPCR (PMA-qPCR), scientists have applied this technology to detect viable cells of various bacterial pathogens in foods. The incorporation of PMA into PCR-based assays for viability detection of pathogens in foods has increased significantly in the last decade. On the other hand, some downsides with this approach have been noted, particularly to achieve complete suppression of signal of DNA from the dead cells present in some particular food matrix. Nowadays, there is a tendency of more and more researchers adapting this approach for viability detection; and a few commercial kits based on PMA are available in the market. As time goes on, more scientists apply this approach to a broader range of pathogen detections, this viability approach (PMA or other chemicals such as platinum compound) may eventually become a common methodology for the rapid, sensitive, and accurate detection of foodborne pathogens. In this review, we summarize the development in the field including progress and challenges and give our perspective in this area.

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“…34 While it is plausible that water contaminated by H. pylori is a route for the transmission of H. pylori infection, the quantities of H. pylori reported to be in drinking water and thus the risk of infection from such sources remain poorly characterized. 37 Both studies had limited sample sizes (n = 87 and n = 24, respectively) and were conducted in a variety of locations. To our knowledge, only two studies have quantitatively measured H. pylori in municipal drinking water: one by our group in Lima, Peru, 26 and the other in Spain.…”

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confidence: 99%

An assessment of drinking water contamination with Helicobacter pylori in Lima, Peru

et al. 2018

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This study has provided evidence of the presence of H. pylori DNA in the drinking water of a single drinking water faucet in the Lince district of Lima. However, no seasonal trends were observed. Further studies are needed to determine the presence of H. pylori in other drinking water sources in other districts in Lima, as well as to determine the viability of H. pylori in these water sources. Such studies would potentially allow for better understanding and estimates of the risk of infection due to exposure to H. pylori in drinking water.

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Identification of Viable Helicobacter pylori in Drinking Water Supplies by Cultural and Molecular Techniques

Abstract: Our results demonstrate for the first time that H. pylori can survive and be potentially infective in drinking water, showing that water distribution systems could be a potential route for H. pylori transmission.

Cited by 73 publications

References 32 publications

Molecular monitoring of disinfection efficacy of E. coli O157:H7 in bottled purified drinking water by quantitative PCR with a novel dye

Molecular monitoring of disinfection efficacy of E. coli O157:H7 in bottled purified drinking water by quantitative PCR with a novel dye

Advances and Challenges in Viability Detection of Foodborne Pathogens

An assessment of drinking water contamination with Helicobacter pylori in Lima, Peru

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