Detection of Helicobacter pylori in sewage and water using a new quantitative PCR method with SYBR® green

Abstract

Aims: To demonstrate the application of a new quantitative polymerase chain reaction (qPCR) technique for the determination of Helicobacter pylori concentrations in water, and to use this method to investigate the occurrence of the bacteria in sewage. The other aim was to study the survival capacity and detectability of the bacteria in artificially contaminated groundwater at different temperatures of 4 and 15°C. Methods and Results: The detection of H. pylori in water was aided by PCR using specific primers designed for the amplification of a fragment within the major vacuolating cytotoxin gene. Conventional culture was compared with conventional PCR and the new real‐time (RT) qPCR approach for the quantification of the bacterium. Helicobacter pylori remained culturable for 120 h at 4°C as opposed to only 24 h at 15°C. RT qPCR demonstrated a 100‐fold greater sensitivity for the detection of H. pylori DNA in comparison with conventional PCR. Scanning electron microscopic (SEM) observation showed that the normal spiral form changed to a coccoid form after 24 and 72 h at 15 and 4°C, respectively. Helicobacter pylori was found at 2–28 cells ml−1 in sewage, of the 23 sewage samples – 84% were positive for H. pylori species‐specific vacuolating cyctotoxin gene (vacA) by RT qPCR, but were negative by conventional PCR. Conclusions: The RT qPCR assay provided a specific, sensitive and rapid method for the quantitative detection of H. pylori in sewage. This molecular method would be valuable in studying the prevalence of H. pylori as required by the United States Environmental Protection Agency Contaminant Candidate List, particularly in nondisinfected ground waters, in sewage as a source of contamination, and for addressing the possible presence of viable but nonculturable of H. pylori. Significance and Impact of the Study: The quantitative detection of H. pylori by rapid and less‐expensive methods than the TaqMan Assay using SYBR green could be an important tool to monitor infection in community by measuring the concentrations in sewage and to meet the new regulatory and risk‐based frameworks for water supplies.

Risk Factors Associated with Helicobacter Pylori Infections in Makurdi Northcentral Nigeria

“…The findings of this study did not also agree with the study by Papiez et al that water-borne transmission of H. pylori could be an important source of infection in developing countries [8]. A study in Guatemala reported that H. pylori was unlikely to be transmitted by water [17] but epidemiological studies in many countries such as China [18], Lima Peru [19], and Colombia [20] had shown that infection was related to H. pylori contaminated water sources. Dube et al also reported that H. pylori could live in tap water in its infectious bacillary form, and in river water for several months in the nonculturable but viable cocoid form.…”

“…Ahmed et al in South India also reported that the prevalence of H. pylori infection among people who drank well water was 92% compared with 74.8% of those who drank tap water (p<0.001) [3]. Several Epidemiological studies in many countries such as rural China, Lima Peru and Colombia had also shown that infection was related to H. pylori contaminated water sources [18][19][20]. However, in agreement with our study, Klein et al reported in a study in Guatemala that H. pylori were unlikely to be transmitted by water [17].…”

Novel Probiotic Bifidobacterium bifidum CECT 7366 Strain Active against the Pathogenic Bacterium Helicobacter pylori

“…In all cases, H. pylori was quantified by real-time PCR as described by Nayak and Rose (39). H. pylori was grown in BHI medium supplemented with 5% (vol/vol) FBS under microaerobic conditions at 37°C for 3 days.…”

Amoeba-Related Health Risk in Drinking Water Systems: Could Monitoring of Amoebae Be a Complementary Approach to Current Quality Control Strategies?

“…Infection with H. pylori is estimated to be 40-70% worldwide [18]. While H. pylori infection sources remain uncertain, recent studies have reported H. pylori DNA detection in water samples [19][20][21][22]. This evidence suggests an extensive distribution of H. pylori in aquatic environments and may indicate water itself as a potential infection source.…”