A combination of direct viable count and fluorescent in situ hybridization for estimating Helicobacter pylori cell viability

Piqueres, Patricia; Moreno, Yolanda; Alonso, J.L.; Ferrús, María Antonia

doi:10.1016/j.resmic.2005.09.003

Cited by 52 publications

(65 citation statements)

References 24 publications

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“…These findings were in line with those of the previous studies of the authors, but further investigation is required to determine whether the organism is viable or not (39). These findings also confirmed the previous observations in Isfahan, Iran (13), Colombia (40), Peru (41), Mexico (42), England (43), Sweden (38,39,41,42,44), Japan (45), and the United States (46). The high prevalence of H. pylori detected in drinking water samples strengthens the evidence of H. pylori transmission through drinking water.…”

Section: Discussionsupporting

confidence: 91%

Detection of Helicobacter pylori in Drinking Water by Loop-Mediated Isothermal Amplification

Abiri

Bagherabadi

Kashef

et al. 2017

Jundishapur J Microbiol

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Background: There should be a public environmental reservoir for Helicobacter pylori in the developing countries, such as Iran, due to their high infection rate of over 70%. Epidemiological findings revealed that water could be a possible source of H. pylori transmission. However, high prevalence of H. pylori in drinking water in Kermanshah, West of Iran, was detected in the authors' previously published study. The current study aims at designing a more accurate and rapid procedure to investigate the prevalence of Helicobacter species and cagA gene in drinking water samples in Kermanshah, from October to December 2012. Methods: In the current study, 60 tap water samples were obtained and specific polymerase chain reaction (PCR) targeted cagA and 16s rRNA was performed. A loop-mediated isothermal amplification (LAMP) targeted ureC gene was developed to accurately detect H. pylori in water samples. Results: The prevalence of ureC by PCR, ureC by LAMP and 16s rRNA by PCR were 26.67%, 38%, and 61.67%, respectively. Among 24 samples (40%), 1 of the 2 tests was positive. The prevalence of cagA gene among ureC positive, 16s rRNA positive and all samples were 18.75%, 13.51%, and 10%, respectively. Conclusions: Helicobacter pylori contamination in drinking water was considerably higher using LAMP compared with PCR. It is noteworthy that some H. pylori positive samples were also positive for Caga.

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Section: Discussionsupporting

confidence: 91%

Detection of Helicobacter pylori in Drinking Water by Loop-Mediated Isothermal Amplification

Abiri

Bagherabadi

Kashef

et al. 2017

Jundishapur J Microbiol

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“…FISH and PCR techniques do not allow discrimination between dead and viable cells Okoh et al, 2007 . A direct viable count FISH procedure DVC-FISH has been proposed for monitoring viable microbial cells in different environments Baudart et al, 2002, Piqueres et al, 2006 . This technique may be useful for detecting and tracing viable phylloepiphytic and endophytic microbial cells in cultivated plants and farming soil.…”

Section: Discussionmentioning

confidence: 99%

Development of a PCR-Based Method for Monitoring the Status of Alcaligenes Species in the Agricultural Environment

Nakano

Niwa²

2014

Biocontrol Sci.

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To analyze the status of the genus Alcaligenes in the agricultural environment, we developed a PCR method for detection of these species from vegetables and farming soil. The selected PCR primers amplified a 107-bp fragment of the 16S rRNA gene in a specific PCR assay with a detection limit of 1.06 pg of pure culture DNA, corresponding to DNA extracted from approximately 23 cells of Alcaligenes faecalis. Meanwhile, PCR primers generated a detectable amount of the amplicon from 2.2 10 2 CFU/ml cell suspensions from the soil. Analysis of vegetable phylloepiphytic and farming soil microbes showed that bacterial species belonging to the genus Alcaligenes were present in the range from 0.9 10 0 CFU per gram or cm 2 Japanese radish: Raphanus sativus var. longipinnatus to more than 1.1 10 4 CFU/g broccoli flowers: Brassica oleracea var. italic , while 2.4 10 2 to 4.4 10 3 CFU/g were detected from all soil samples. These results indicated that Alcaligenes species are present in the phytosphere at levels 10-1000 times lower than those in soil. Our approach may be useful for tracking or quantifying species of the genus Alcaligenes in the agricultural environment.

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“…Since its first description by Kogure et al (17), the DVC method has been used widely in microbial ecology, mostly for aquatic samples including seawater (14,17,18), freshwater (3,4,16,24), groundwater (2), wastewater (8), and drinking water (4,5,11). Its application to terrestrial environments such as soil and compost, however, is very limited.…”

Section: Discussionmentioning

confidence: 99%

Direct Viable Count Combined with Fluorescence in situ Hybridization (DVC-FISH) for Specific Enumeration of Viable Escherichia coli in Cow Manure

Ueno

Inoue

et al. 2009

Microb. Environ.

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A direct viable count procedure combined with fluorescence in situ hybridization (DVC-FISH) was developed for the specific detection and enumeration of viable Escherichia coli in cow manure. The DVC method was performed by trapping bacterial cells, extracted from cow manure samples, onto Nucleopore filters followed by incubation on a DVC medium containing yeast extract and four gyrase inhibitors. E. coli cells were identified by using the probe ES445. The DVC method efficiently promoted the elongation of E. coli cells and allowed for the recognition of individual cell division events, by observing microcolonies. Cell recovery by DVC-FISH together with bacterial extraction, was 53% with an inoculum of 10 7 to 10 10 cells g −1 dry weight, when the manure samples were inoculated with a fresh culture of E. coli and determinations were made immediately. An examination of the survival of E. coli in a cow manure microcosm showed that an increasing fraction of E. coli became non-culturable but were still detectable by DVC-FISH. All these results suggest that DVC-FISH is useful for enumerating viable, even non-culturable, E. coli in cow manure.

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A combination of direct viable count and fluorescent in situ hybridization for estimating Helicobacter pylori cell viability

Cited by 52 publications

References 24 publications

Detection of Helicobacter pylori in Drinking Water by Loop-Mediated Isothermal Amplification

Detection of Helicobacter pylori in Drinking Water by Loop-Mediated Isothermal Amplification

Development of a PCR-Based Method for Monitoring the Status of Alcaligenes Species in the Agricultural Environment

Direct Viable Count Combined with Fluorescence in situ Hybridization (DVC-FISH) for Specific Enumeration of Viable Escherichia coli in Cow Manure

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