Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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PCR-Based RELP Analysis of ureC Gene for Typing of Indian Helicobacter pylori Strains from Gastric Biopsy Specimens and Culture

  • Mishra, Kanchan-Kumar (Department of microBiology Sanjay Gandhi Postgraduate Institute of Medical Sciences) ;
  • Prabhat P. Dwivedi (Department of microBiology Sanjay Gandhi Postgraduate Institute of Medical Sciences) ;
  • Prasad, Kashi-Nath (Department of microBiology Sanjay Gandhi Postgraduate Institute of Medical Sciences) ;
  • Archana Ayyagari (Department of microBiology Sanjay Gandhi Postgraduate Institute of Medical Sciences)
  • Published : 2002.12.01
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Abstract

Since culture of Helicobacter pylori is relatively insensitive and cumbersome, molecular detection and typing of H. pylori isolates are gaining importance for strain differentiation. In the present study genomic DNA of 42 gastric biopsies and H. pylori isolates from corresponding patients were analyzed and compared by PCR-based RFLP assay. The 1,132-bp product representing an internal portion of ureC gene of H. pylori was amplified by PCR and digested with restriction enzymes HindⅢ, AiuⅠ and PvuⅠ. The HindⅢ, AluⅠ and PvuⅠ digestion produced 4, 7, and 2 distinguishable RFLP patterns respectively from 42-H. pylori isolates. By combining all three restriction enzyme digestions, 15 RFLP patterns were observed. However, when PCR products from 42 gastric biopsy specimens were digested by restriction enzymes HindⅢ, AluⅠ and PvuⅠ, we observed 5, 8 and 2 RFLP patterns, respectively. Patterns from 34 of 42 gastric biopsy specimens matched those of corresponding H. pylori isolates from respective patients. Patterns from the remaining eight biopsy specimens differed and appeared to represent infection with two H. pylori strains. The patterns of one strain from each of these biopsies was identical to that of the isolate from corresponding patients and the second pattern presumably represented the co-infecting strain. From the study, it appears that PCR-based RFLP analysis is a useful primary tool to detect and is distinguish H. pylori strains from gastric biopsy specimens and is superior to culture techniques in the diagnosis of infection with multiple strains of H. pylori.

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