To set up a method of amplification for the whole CagA gene of Helicobacter pylori and its fingerprinting by restriction fragment length polymorphism(RFLP),nested PCR was employed in combination with TD-PCR to amplify the gene and EcoRI and Hind Ⅲ were used to generate the RFLP fingerprinting.Target DNA fragments from 13 of 20 samples were successfully amplified and the relevant RFLP fingerprintings were obtained.It is concluded that the method can be used to amplify the whole CagA gene and CagA gene has apparent diversity of RFLP profile. 相似文献
ObjectiveTo investigate the prevalence of Helicobacter pylori (H. pylori) infection in dyspepsia patients and its relation to virulence factor cagA gene.MethodsIn total, 110 gastric biopsies from dyspeptic patients were comparatively studied using rapid urease test and multiplex PCR. Multiplex PCR detected three genes of 16S rRNA, cagA, and ureC.ResultsH. pylori was detected in 14 gastric biopsies (13%). Significantly higher number of female were infected. Furthermore, cagA gene was found in all H. pylori–positive specimens. In addition, the result indicated that the multiplex PCR with annealing temperature at 57 oC was able to effectively amplify specific products.ConclusionThe results confirmed that high preva¬lence of cagA gene in H. pylori among dyspeptic patients in Southern Thailand. 相似文献
a Institute of Internal Medicine, University of Siena, Siena, Italy, b Institute of Pathology, University of Siena, c IRIS, Siena, d Institute of Surgical Clinics, University of Siena, e Institute of Internal Medicine, University of Bologna, Bologna, Italy
Correspondence to: Dr N Figura, Institute of Internal Medicine, University of Siena, Policlinico Le Scotte, viale Bracci, I-53100 Siena, Italy.
Accepted for publication 6 February 1998
Background/Aims—Infection with Helicobacter pylori strains harbouring the cagA gene (cagA+) is associated with an increased risk of developing peptic ulcer and gastric cancer. The aim of this study was to assess whether H pylori isolates with different cagA status were present in patients with non-ulcer dyspepsia, and whether a variable cagA status is relevant to histological gastric mucosal damage and glandular cell proliferation. Methods—Well separated H pylori colonies (between 2 and 25) from primary plates, per gastric area, for each of 19 patients with non-ulcer dyspepsia were examined for cagA by hybridisation. Western blotting was used to examine both representative colonies for CagA expression and the patients' sera for antibody response to CagA. Glandular gastric cell proliferation was assessed immunohistochemically. Results—Of the 747 colonies examined, 45.3% were cagA+. All colonies from four patients were cagA+, and all colonies from two patients were cagA−. In 13 patients (68%) both cagA+ and cagA− colonies were found. CagA expression of isolates corresponded to their cagA status. H pylori strains with different CagA molecular masses were present in three patients. Results based on all 19patients studied showed that the prevalence of cagA+ colonies in areas with mucosal atrophy associated or not with intestinal metaplasia (67.9%) was significantly higher than in normal mucosa (44.7%) and mucosa from patients with chronic gastritis (44.0%) (p< 0.001). High levels of cell proliferation were associated with histological atrophy with or without intestinal metaplasia, but not with the possession of cagA by organisms colonising the same mucosal sites. Conclusions—Most patients with non-ulcer dyspepsia are infected by both cagA+ and cagA− H pylori colonies. The cagA status of infecting organisms may play a role in the development of atrophy and intestinal metaplasia. (GUT 1998;:772-778)