Rapid and sensitive detection of infectious pathogens like H. pylori and C. jejuni is necessary for the maintenance of a safe food or water supply. The first portion of this research aimed to demonstrate the application of a new qPCR technique for determination of H. 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 4AdC and 15AdC. Real Time qPCR demonstrated a 100-fold greater sensitivity for detection of H. pylori DNA in comparison to conventional PCR. SEM observation showed that the normal spiral form changed to a coccoid form after 24hr and 72hr at 15AdC and 4AdC, respectively. H. pylori was found at 2 to 38 cells ml-1 in sewage and 64% (n=39) were positive for H. pylori species specific gene vacA by real time qPCR. The second portions of this research aimed to survey and quantify C. jejuni in sewage using real time q PCR. Twenty-two samples were analyzed and 59% of the sewage samples were positive for the C. jejuni specific gene of gyrA by real time qPCR. Real time qPCR assay provided a specific, sensitive and rapid method for studying the prevalence of H. pylori and C. jejuni and can be used as required by the US EPA Contaminant Candidate List for survey. This study demonstrates that untreated sewage is a source of these bacteria and has the potential to contaminate other waters.After that, the purified target of part of gyrA gene was cloned into ... 2.1 TOPOAr) and transformed into TOP 10 Competent cells (One ShotAr Chemical Transformation) based on TOPO TA Cloning Instruction Manual (Invitrogen, Carlsbad, CA).
|Title||:||Stability and Quantitative Surveillance of Helicobacter Pylori and Campylobacter Jejuni in Environmental Waters by Real Time QPCR.|
|Author||:||Arun Kumar Nayak|
|Publisher||:||ProQuest - 2008|