Sensitivity of culture compared with that of polymerase chain reaction for detection of Helicobacter pylori from antral biopsy samples

Zwet, A. A. Van; Thijs, J. C.; Kooistra-Smid, A.M.D.; Schirm, J; Snijder, J.

doi:10.1128/jcm.31.7.1918-1920.1993

Cited by 69 publications

(26 citation statements)

References 11 publications

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“…In HP-HJM 12 and 14, no positive hybridisation signal was detected, despite the fact that a weak genomic cagA PCR amplicon was visualised. Moreover, hybridisation analysis revealed duplex cagA amplicons (different sizes) derived from cDNA which were not present in genomic DNA from HP- HJM 5,11,13,17,18,22, and 23, respectively [ Table 2].…”

Section: Resultsmentioning

confidence: 99%

“…M olecular typing methods such as ribotyping [1] and randomly amplified polymorphic DNA (RAPD) analysis [2] have demonstrated considerable genomic diversity among Helicobacter pylori isolates. During the past few years, numerous PCR assays have been developed for the identification of specific virulence genes in H. pylori including ureA [3][4][5][6], ureC [7,8], cagA [3,4,9], vacA [10][11][12][13][14][15][16] and the 16S rRNA genes [17,18]. Recently, we have described a method for the rapid simultaneous molecular identification and subtyping of H. pylori by pyrosequencing of the 16S-rDNA variable V1 and V3 region [19].…”

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confidence: 99%

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Molecular Typing of Helicobacter pylori by Virulence‐Gene Based Multiplex PCR and RT‐PCR Analysis

Monstein

Ellnebo-Svedlund²

2002

Helicobacter

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Section: Resultsmentioning

confidence: 99%

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Molecular Typing of Helicobacter pylori by Virulence‐Gene Based Multiplex PCR and RT‐PCR Analysis

Monstein

Ellnebo-Svedlund²

2002

Helicobacter

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“…Although the moderately low OD 450 s of specimens from these patients (only a few units above the cutoff for one of them) could suggest false-positive results, the presence of chronic active gastritis (12) in both patients and the precautions taken to avoid sample contamination suggest that H. pylori has not been eradicated from these patients. The reduction in the number of bacteria and the presence of coccoid forms induced by the antimicrobial agents could account for the failure of the culture to detect H. pylori shortly after the end of treatment (4,22), although culture has been shown in one study (32) to be as sensitive as PCR 3 months after treatment. Furthermore, RUT and histological examination have been described to have decreased sensitivities when used in treatment follow-up (21,25).…”

Section: Discussionmentioning

confidence: 99%

Rapid colorimetric hybridization assay for detecting amplified Helicobacter pylori DNA in gastric biopsy specimens

et al. 1996

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A very simple, practical, sensitive, and specific colorimetric hybridization assay for detecting amplified Helicobacter pylori DNA is described. This assay, which combines a sensitive sandwich DNA hybridization reaction and a colorimetric protocol similar to those used in conventional enzyme immunoassays, was shown to be suitable for detecting H. pylori-infected gastric biopsy specimens and for monitoring the eradication of the pathogen after treatment. The specificity and sensitivity of the colorimetric hybridization assay were tested by assaying 27 H. pylori strains (4 reference and 23 clinical isolates), 9 strains of other Helicobacter spp. or Campylobacter spp., and 11 clinical isolates of other urease-positive bacteria. The likelihood of H. pylori detection in gastric biopsy specimens by the colorimetric hybridization assay was evaluated with 23 H. pylori-positive and 41 H. pylori-negative biopsy specimens on the basis of positive and negative results, respectively, of culture, rapid urease test, histological examination, and PCR. Biopsy specimens from 33 treated patients, endoscopied 4 to 8 weeks after the end of treatment, were also tested. All H. pylori strains showed positive results in the colorimetric hybridization assay, presenting optical densities at 450 nm (OD 450 s) of >3.0. None of the other Helicobacter spp., Campylobacter spp., or the clinical isolates of other urease-positive bacteria showed OD 450 s equal to or greater than the cutoff (mean OD 450 cutoff, 0.208). The colorimetric hybridization assay detected all 23 H. pylori-positive biopsy specimens (mean OD 450 , 2.910 ؎ 0.295), while none of the H. pylori-negative biopsy specimens was shown to be positive in the assay (mean OD 450 , 0.108 ؎ 0.025). H. pylori was considered to be not eradicated from three of the posttreatment biopsy specimens by culture, rapid urease test, histological examination, and PCR. They were all positive by the colorimetric hybridization assay, and their OD 450 s were >3.0. The colorimetric hybridization assay also detected two other H. pyloripositive patients. Specimens from these two patients had negative culture, rapid urease test, and histology results, and a specimen from one of them also tested negative by PCR. These results indicate that the colorimetric hybridization assay is a suitable method both for the diagnosis of H. pylori in biopsy specimens and for the follow-up of patients after the end of treatment.

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“…Polymerase;8 pL MgCl2 (2.0 mM); 2 mM of HP1 and HP2; and 5 pL of DNA template. The mixture was made up to 100 pL by adding ultrapure water, which was then topped with 50 pL sterile mineral oil to prevent evaporation during temperature cycling.A 2-step PCR reaction cycle was used in an automated thermocyler.…”

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Helicobacter pylori: The Mouth, Stomach, and Gut Axis

et al. 1998

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The aim of this study was to identify the natural reservoir and route of transmission of Helicobacter pylori infection. Two hundred eight (208) dyspeptic patients (114 males, 94 females; peak age of cohort, 50-59.9) were recruited. Specimens were collected from saliva, supra- and subgingival dental plaque, tongue scrapings, and oropharyngeal swabs. At subsequent endoscopy, gastric antral biopsy was performed for the rapid urease test (RUT), microbiological culture, and, in some patients, histology. Gastric juice samples were aspirated, and in 50 patients duodenal aspirate was collected. Polymerase chain reaction (PCR) with primers targeted to the 16S rRNA sequence of H. pylori was also employed for each of the specimens. In those patients where H. pylori was detected from multiple sites (dental plaque, gastric juice, gastric biopsy, and duodenal aspirate), restriction endonuclease digestion with Hae III was performed to determine if they were epidemiologically linked. The results indicated that 15/208 patients (7%) tested positively for H. pylori by PCR in dental plaque; only 2 samples were positive by culture. In none of the other oral sites sampled was H. pylori detected by any test used in the study. Gastric juice and gastric biopsy specimens from 36/ 208 patients (17%) and 114/208 patients (55%), respectively, were positive by PCR. Duodenal aspirate from 6/50 patients (12%) also tested positively by PCR. All specimens tested by restriction endonuclease digestion with Hae III (15/15 patients) were positive in both antral biopsy and gastric juice specimens, as well as 5 specimens from the duodenal aspirate. Four of the dental plaque strains had restriction patterns similar to those of the stomach and duodenal sites, providing evidence that these sites were infected with the same strain of H. pylori. In conclusion, the results suggest that H. pylori selects the gastric mucosa as its preferred site. The detection in dental plaque could indicate that the oral cavity may act as a reservoir or sanctuary for the organism. Whether H. pylori is a resident or transient oral microorganism is still unclear, although it is more likely to be transient in nature.

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Sensitivity of culture compared with that of polymerase chain reaction for detection of Helicobacter pylori from antral biopsy samples

Cited by 69 publications

References 11 publications

Molecular Typing of Helicobacter pylori by Virulence‐Gene Based Multiplex PCR and RT‐PCR Analysis

Molecular Typing of Helicobacter pylori by Virulence‐Gene Based Multiplex PCR and RT‐PCR Analysis

Rapid colorimetric hybridization assay for detecting amplified Helicobacter pylori DNA in gastric biopsy specimens

Helicobacter pylori: The Mouth, Stomach, and Gut Axis

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