Direct detection and amplification of Helicobacter pylori ribosomal 16S gene segments from gastric endoscopic biopsies

Hoshina, Sadayori; Kahn, Scott M.; Jian, Wei; Green, Peter H.R.; Neu, Harold C.; Chin, Nai-Xun; Morotomi, Masami; LoGerfo, Paul; Weinstein, I. Bernard

doi:10.1016/0732-8893(90)90079-b

Cited by 71 publications

(51 citation statements)

References 14 publications

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“…Culture and histological examination of biopsy specimens using different stains and assaying for urease activity have the disadvantages of lack of sensitivity and long incubation periods. Assays based on the use of PCR to detect the presence of H. pylori DNA using several different gene targets showed that PCR is feasibile for the rapid, sensitive and specific detection of H. pylori (2)(3)(4)(5)(7)(8)(9)(10)12). using the Pyromark iD system with SQA software and the SQA reagent kit to amplify a section of the 16S rRNA gene it was possible to analyze bacterial genetic targets in DNA extracted directly from human gastric tissues without the prolonged culturing of bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…H. pylori infection in gastric specimens can be demonstrated through the use of culture, histological examination of biopsy specimens using different stains, assaying for urease activity and PCR assay with the aim of specifically detecting H. pylori DNA (2). Assays based on the use of PCR to detect the presence of H. pylori DNA using several different gene targets have been described (2)(3)(4)(5)(6)(7)(8)(9)(10). Moreover, it is well known that the PCR assay is highly reliable in the detection of H. pylori.…”

Section: Introductionmentioning

confidence: 99%

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Molecular identification of Helicobacter DNA in human gastric adenocarcinoma tissues using Helicobacter species-specific 16S rRNA PCR amplification and pyrosequencing analysis

Han

Lee²,

Lim³

et al. 2010

Oncology Letters

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Abstract.Helicobacter pylori (H. pylori) is a microaerophilic gram-negative bacterium known to be associated with chronic gastritis, peptic ulcer and gastric adenocarcinoma. In the present study, the presence of Helicobacter DNA was investigated using a Helicobacter species-specific 16S rRNA PCR amplification and pyrosequencing analysis in 51 resected gastric adenocarcinomas. DNA was extracted from paraffinembedded tissues of resected gastric adenocarcinomas. PCR primers were designed to amplify the 133-bp PCR fragment in highly conserved regions of the 16S rRNA gene. The sequence of the PCR products was analyzed using a PSQ 96 system with SQA software. The pyrosequencing analysis of 16S rRNA showed that H. pylori was present in 47 (92.2%) of the 51 gastric adenocarcinomas. In the 4 H. pylori-negative cases, Helicobacter cinaedi (2 cases), Helicobacter mustelae (1 case) and Campylobacter hyointestinalis (1 case) were detected. Pyrosequencing technology was useful in the identification and differentiation of H. pylori from other species by analyzing the gene encoding 16S rRNA. Gastric adenocarcinoma tissues contain bacteria, and the majority are H. pylori. Helicobacter cinaedi, Helicobacter mustelae and Campylobacter hyointestinalis rarely occur. The roles of these organisms in the pathogenesis of gastric adenocarcinoma remain unclear.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular identification of Helicobacter DNA in human gastric adenocarcinoma tissues using Helicobacter species-specific 16S rRNA PCR amplification and pyrosequencing analysis

Han

Lee²,

Lim³

et al. 2010

Oncology Letters

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“…The current gold standard for diagnosing H. pylori infection is endoscopic biopsy of gastric tissue for the rapid urease test, histology, and culture. However, such an invasive procedure has major disadvantages of anesthesia, discomfort, and the possibility for ethical problems (Hoshina et al, 1990). But, noninvasive tests are easy to perform and do not produce significant discomfort and allow a patient to avoid the discomfort and risk of invasive endoscopy.…”

Section: Discussionmentioning

confidence: 99%

“…But, noninvasive tests are easy to perform and do not produce significant discomfort and allow a patient to avoid the discomfort and risk of invasive endoscopy. These include serological antibody testing for H. pylori, the urea breath test, and the stool antigen assay (HpSA) test (Hoshina et al, 1990).…”

Section: Discussionmentioning

confidence: 99%

Comparison of the non-invasive diagnostic methods, stool antigen test and PCR assay, for Helicobacter felis detection in dogs

Hong¹,

Lee²,

Kim³

et al. 2015

Korean Journal of Veterinary Service

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The aim of the present study was to compare the non-invasive methods for the diagnosis of H. felis with HpSA kit-based detection method and H. felis-specific PCR assay with dog's stool samples without sacrifice. Male Beagle dogs (n=6) were infected with H. felis ATCC 49179 (1.0×10 9 CFU/dog) by intragastric inoculation two times at 3-day intervals, and the stool specimens of dogs were collected 1, 3, 5, 7, 14, 21 days after infection to submit to HpSA test and H. felis-specific PCR. As the results, the sensitivity of the HpSA and the PCR analysis was 50.0%, 83.3% respectively. Although HpSA test is less sensitive, it could be used for rapid, cheap and easy screening assay for H. felis infection in dog and cats. We suggest that the H. pylori stool antigen kit, HpSA, is useful and effective for monitoring H. felis infection. If HpSA test would be made with H. felis antibodies in the future, its sensitivity could be increased. Also, PCR assay could be successfully used to detect the H. felis in stools. Applying the H. pylori stool antigen kit and PCR assay may be the recommended non-invasive strategy to identify H. felis in dog and cats.

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“…The fragments of partial 16S rDNAs from TA-1, TA-2, N-8, A. infernus and D, ambivalens were amplified by PCR with synthetic oligonucleotides (5'-CAG CAG CCG CGG TAA TAC and 5'-ACG GGC GGT GTG TGC). These sequences are universally found around positions 530 and 1400, respectively in the 16S rRNA genes not only in eubacteria but also in archaebacteria (7). The amplified fragments were subcloned into pBluescript SK-(Stratagene, La Jolla, U.S.A.).…”

mentioning

confidence: 99%

Characterization and identification of thermoacidophilic archaebacteria isolated in Japan.

Kurosawa

Sugai

Fukuda

et al. 1995

J. Gen. Appl. Microbiol.

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Three isolates of spherical thermoacidophilic archaebacteria (strains TA-1, TA-2 and N-8) were obtained from acidic hot springs at Hakone and Noboribetsu in Japan. We have studied the isolates using electron microscopy, Gram staining, sensitivity to lysozyme, growth conditions, lipid compositions, SDS-PAGE patterns of whole-cell proteins, DNA base composition and partial 16S rRNA sequences. These characteristics of the isolates were compared with those of the type strains, Sulfolobus acidocaldarius, Sulfolobus solfataricus, Acidianus brierleyi, Acidianus infernus, Metallosphaera sedula and Desulfurolobus ambivalens. Comparative studies indicated that the isolate TA-1 was novel species of either Sulfolobus or Acidianus, the isolate TA-2 belonged to the genus Metallosphaera, and the isolate N-8 belonged to the same species of S. acidocaldarius isolated from Yellowstone.A comparative sequence study of the 16S rRNAs showed that living organisms can be divided into three kingdoms: the traditional eubacterial and eukaryotic kingdoms and the newly recognized archaebacterial kingdom (22,23). Although archaebacteria exhibit a prokaryotic cell structure and organization, they show distinguishing and unique features (1,10,11,(14)(15)(16).From acidic hot springs in Japan, three isolates of spherical thermoacidophilic archaebacteria (strains TA-1, TA-2 and N-8) were obtained. Total lipid composition analysis and preliminary results of growth temperature and pH showed that these three strains belong to the order Sulfolobales (19,20).

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Direct detection and amplification of Helicobacter pylori ribosomal 16S gene segments from gastric endoscopic biopsies

Cited by 71 publications

References 14 publications

Molecular identification of Helicobacter DNA in human gastric adenocarcinoma tissues using Helicobacter species-specific 16S rRNA PCR amplification and pyrosequencing analysis

Molecular identification of Helicobacter DNA in human gastric adenocarcinoma tissues using Helicobacter species-specific 16S rRNA PCR amplification and pyrosequencing analysis

Comparison of the non-invasive diagnostic methods, stool antigen test and PCR assay, for Helicobacter felis detection in dogs

Characterization and identification of thermoacidophilic archaebacteria isolated in Japan.

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