Usefulness of Housekeeping Genes for the Diagnosis of Helicobacter pylori Infection, Strain Discrimination and Detection of Multiple Infection

Abstract

<b>Background:</b> <i>Helicobacter pylori</i> infects human stomachs of over half the world's population, evades the immune response and establishes a chronic infection. Although most people remains asymptomatic, duodenal and gastric ulcers, MALT lymphoma and progression to gastric cancer could be developed. Several virulence factors such as flagella, lipopolysaccharide, adhesins and especially the vacuolating cytotoxin VacA and the oncoprotein CagA have been described for <i>H. pylori</i>. Despite the extensive published data on <i>H. pylori</i>, more research is needed to determine new virulence markers, the exact mode of transmission or the role of multiple infection. <b>Materials and Methods:</b> Amplification and sequencing of six housekeeping genes (<i>amiA</i>, <i>cgt</i>, <i>cpn60</i>, <i>cpn70</i>, <i>dnaJ</i>, and <i>luxS</i>) related to <i>H. pylori</i> pathogenesis have been performed in order to evaluate their usefulness for the specific detection of <i>H. pylori</i>, the genetic discrimination at strain level and the detection of multiple infection. A total of 52 <i>H. pylori</i> clones, isolated from 14 gastric biopsies from 11 patients, were analyzed for this purpose. <b>Results:</b> All genes were specifically amplified for <i>H. pylori</i> and all clones isolated from different patients were discriminated, with gene distances ranged from 0.9 to 7.8%. Although most clones isolated from the same patient showed identical gene sequences, an event of multiple infection was detected in all the genes and microevolution events were showed for <i>amiA</i> and <i>cpn60</i> genes. <b>Conclusions:</b> These results suggested that housekeeping genes could be useful for <i>H. pylori</i> detection and to elucidate the mode of transmission and the relevance of the multiple infection.