Quantification of Helicobacter pylori in the viable but nonculturable state by quantitative PCR in water disinfected with ozone

Water Sci Technol. 2013;68(11):2468-72. doi: 10.2166/wst.2013.512.

Abstract

Helicobacter pylori is a Gram-negative spiral-shaped bacterium that colonizes the gastric mucosa and is associated with gastric diseases. It may present a morphological adaptation when it is out of its natural environment, such as in water. The morphological adaptation is a coccoid form, which is a viable but non-culturable state (VNC) in which the DNA remains active and therefore infective. Due to the impossibility of culture by traditional methods in the VNC state, we developed a methodology that includes a molecular technique, quantitative polymerase chain reaction (qPCR), which is capable of measuring the bacteria in both forms (helical and coccoidal) and therefore is able to measure a disinfection process and to estimate the resistance of the bacteria to ozone. The methodology developed measures the efficiency of the ozone disinfection when bacteria are in a VNC state only. Bacterial culture at 9 × 10(8)CFU/mL diluted in 40 mL reaction volumes were exposed to a wide range of CT values (0.11-15 mg min/L). The results show a 3.92-log reduction when treated with 15 mg min/L. Our results demonstrate the feasibility of using qPCR for the quantification and detection of H. pylori, in coccoid form, in water systems treated with an ozone disinfection process.

Publication types

  • Validation Study

MeSH terms

  • Disinfection
  • Helicobacter pylori / isolation & purification*
  • Ozone
  • Real-Time Polymerase Chain Reaction
  • Water Microbiology*

Substances

  • Ozone