Molecular Detection of Helicobacter pylori in Drinking Water in Khartoum State, 2019

Abstract

Helicobacter pylori (H. pylori) is one of most the causative agent of chronic bacterial infection in humans, and act as predisposing factor for peptic ulcer and gastric cancer. The infection has strongly association with lack of access to clean water and proper sanitation, However H. pylori loses its ability to survive in an infectious state in the environment because it rapidly loses its cultivability. The aim of this study was to detect H. pylori in water using culture and molecular methods. One hundred water samples were collected from tap water with and without filters, cooler and Zeer from different area in Khartoum state. Samples were filtered through 0.45µm filter membrane (cellulose membrane filter). Each membrane was takensliced and immersed in 2 ml of Brain Heart infusion broth (BHI) media (Himedia, India) for overnight. After that each 2 ml of BHI was taken and cultured for H. pylori on special Columbia media (Himedia, India) contain special selective supplement and incubated in closed jars with special kits to provide environment with the oxygen tension lowered to 5-15% and carbon dioxide raised to 1-10% at temperature 37ºC for three days and incubated for a week before being discarded as negative, the identification was depending on their colonial morphology. DNA was extracted by quinidine chloride method from 2 ml Brain Heart Infusion (BHI) broth media and PCR technique were applied to these samples to detect H.pylori genes (16sRNA specific for Helicobacter pylori and Urease C). Out of 100 samples in cultural method there was no growth, and in molecular methodthere was no positive sample for both urease C and 16sRNA genes (0%), This finding indicates that water may not act as source of transmission for H. pylori infection, or may be due to the addition chlorine to water system in Khartoum.