PCR for identification and typing of Helicobacter pylori isolated from children
Dzierzanowska, D.; Gzyl, A.; Rozynek, E.; Augustynowicz, E.; Wojda, U.; Celińska-Cedro, D.; Sankowska, M.; Wadström, T.
Journal of Physiology and Pharmacology An Official Journal of the Polish Physiological Society 47(1): 101-114
1996
ISSN/ISBN: 0867-5910 PMID: 8777290 Document Number: 459647
From 191 children with upper gastroduodenal disease 236 gastric biopsy specimens were taken. H. pylori was detected by use of culture Gram staining histological examination and PCR technique. A segment of DNA coding protein synthesis of 26 kDa or urease A and B gene were used for PCR amplification. PCR technique was also used for determination of the presence of Cag A gene in 72 strains of H. pylori isolated from children. Genetic typing of H. pylori strains by RFLP analysis of PCR amplified urease B gene 933 bp fragment and RAPD were performed. Biopsy specimens taken from children with gastritis were in 52% H. pylori culture and PCR positive, while 18.1% PCR positive only. Similarly, specimens taken from children with duodenal ulcer were in 50% H. pylori culture and PCR positive, while 12.5% PCR positive only. Fifty one (70.8%) from 72 strains of H. pylori were Cag A positive. Molecular typing of the strains isolated during first and follow-up endoscopy allowed the differentiation between reinfection or new infection and coinfection. It was shown that RAPD typing had better discrimination power in comparison to PCR--RFLP method.