Tags

Type your tag names separated by a space and hit enter

Absence of Helicobacter pylori high tetracycline resistant 16S rDNA AGA926-928TTC genotype in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil.
BMC Gastroenterol. 2012 May 17; 12:49.BG

Abstract

BACKGROUND

Treatment effectiveness of Helicobacter pylori varies regionally and is decreasing worldwide, principally as a result of antibiotic resistant bacterium. Tetracycline is generally included in second line H. pylori eradication regimens. In Brazil, a high level of tetracycline resistance (TetR) is mainly associated with AGA926-928TTC 16 S rDNA nucleotide substitutions. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori 16 S rDNA high level TetR genotype using a molecular approach directly on gastric biopsies of dyspeptic patients attending consecutively at Hospital das Clinicas of Marilia, São Paulo, Brazil.

METHODS

Gastric biopsy specimens of 68 peptic ulcer disease (PUD) and 327 chronic gastritis (CG) patients with a positive histological diagnosis of H. pylori were investigated for TetR 16 S rDNA genotype through a molecular assay based on amplification of a 16 S rDNA 545 bp fragment by polymerase chain reaction and HinfI restriction fragment length polymorphism (PCR/RFLP). Through this assay, AGA926-928TTC 16 S rDNA TetR genotype resulted in a three DNA fragment restriction pattern (281, 227 and 37 bp) and its absence originated two DNA fragments (264 and 281 bp) due to a 16 S rDNA conserved Hinf I restriction site.

RESULTS

The 545 bp 16 S rDNA PCR fragment was amplified from 90% of gastric biopsies from histological H. pylori positive patients. HinfI RFLP revealed absence of the AGA926-928TTC H. pylori genotype and PCR products of two patients showed absence of the conserved 16 S rDNA HinfI restriction site. BLASTN sequence analysis of four amplicons (two conserved and two with an unpredicted HinfI restriction pattern) revealed a 99% homology to H. pylori 16 S rDNA from African, North and South American bacterial isolates. A nucleotide substitution abolished the conserved HinfI restriction site in the two PCR fragments with unpredicted HinfI RFLP, resulting in an EcoRI restriction site.

CONCLUSIONS

H. pylori AGA926-928TTC 16 S rDNA gene substitutions were not found in our population. More research is required to investigate if H. pylori TetR has a different genetic background in our region and if the nucleotide substitutions of the uncultured H. pylori 16 S rRNA partial sequences have biological significance.

Authors+Show Affiliations

Department of Genotyping, Hemocenter, Marilia Medical School, Marilia, SP, Brazil.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22594560

Citation

Suzuki, Rodrigo Buzinaro, et al. "Absence of Helicobacter Pylori High Tetracycline Resistant 16S rDNA AGA926-928TTC Genotype in Gastric Biopsy Specimens From Dyspeptic Patients of a City in the Interior of São Paulo, Brazil." BMC Gastroenterology, vol. 12, 2012, p. 49.
Suzuki RB, Almeida CM, Sperança MA. Absence of Helicobacter pylori high tetracycline resistant 16S rDNA AGA926-928TTC genotype in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil. BMC Gastroenterol. 2012;12:49.
Suzuki, R. B., Almeida, C. M., & Sperança, M. A. (2012). Absence of Helicobacter pylori high tetracycline resistant 16S rDNA AGA926-928TTC genotype in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil. BMC Gastroenterology, 12, 49.
Suzuki RB, Almeida CM, Sperança MA. Absence of Helicobacter Pylori High Tetracycline Resistant 16S rDNA AGA926-928TTC Genotype in Gastric Biopsy Specimens From Dyspeptic Patients of a City in the Interior of São Paulo, Brazil. BMC Gastroenterol. 2012 May 17;12:49. PubMed PMID: 22594560.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Absence of Helicobacter pylori high tetracycline resistant 16S rDNA AGA926-928TTC genotype in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil. AU - Suzuki,Rodrigo Buzinaro, AU - Almeida,Cristiane Maria, AU - Sperança,Márcia Aparecida, Y1 - 2012/05/17/ PY - 2012/01/05/received PY - 2012/05/07/accepted PY - 2012/5/19/entrez PY - 2012/5/19/pubmed PY - 2013/3/26/medline SP - 49 EP - 49 JF - BMC gastroenterology JO - BMC Gastroenterol VL - 12 N2 - BACKGROUND: Treatment effectiveness of Helicobacter pylori varies regionally and is decreasing worldwide, principally as a result of antibiotic resistant bacterium. Tetracycline is generally included in second line H. pylori eradication regimens. In Brazil, a high level of tetracycline resistance (TetR) is mainly associated with AGA926-928TTC 16 S rDNA nucleotide substitutions. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori 16 S rDNA high level TetR genotype using a molecular approach directly on gastric biopsies of dyspeptic patients attending consecutively at Hospital das Clinicas of Marilia, São Paulo, Brazil. METHODS: Gastric biopsy specimens of 68 peptic ulcer disease (PUD) and 327 chronic gastritis (CG) patients with a positive histological diagnosis of H. pylori were investigated for TetR 16 S rDNA genotype through a molecular assay based on amplification of a 16 S rDNA 545 bp fragment by polymerase chain reaction and HinfI restriction fragment length polymorphism (PCR/RFLP). Through this assay, AGA926-928TTC 16 S rDNA TetR genotype resulted in a three DNA fragment restriction pattern (281, 227 and 37 bp) and its absence originated two DNA fragments (264 and 281 bp) due to a 16 S rDNA conserved Hinf I restriction site. RESULTS: The 545 bp 16 S rDNA PCR fragment was amplified from 90% of gastric biopsies from histological H. pylori positive patients. HinfI RFLP revealed absence of the AGA926-928TTC H. pylori genotype and PCR products of two patients showed absence of the conserved 16 S rDNA HinfI restriction site. BLASTN sequence analysis of four amplicons (two conserved and two with an unpredicted HinfI restriction pattern) revealed a 99% homology to H. pylori 16 S rDNA from African, North and South American bacterial isolates. A nucleotide substitution abolished the conserved HinfI restriction site in the two PCR fragments with unpredicted HinfI RFLP, resulting in an EcoRI restriction site. CONCLUSIONS: H. pylori AGA926-928TTC 16 S rDNA gene substitutions were not found in our population. More research is required to investigate if H. pylori TetR has a different genetic background in our region and if the nucleotide substitutions of the uncultured H. pylori 16 S rRNA partial sequences have biological significance. SN - 1471-230X UR - https://www.unboundmedicine.com/medline/citation/22594560/Absence_of_Helicobacter_pylori_high_tetracycline_resistant_16S_rDNA_AGA926_928TTC_genotype_in_gastric_biopsy_specimens_from_dyspeptic_patients_of_a_city_in_the_interior_of_São_Paulo_Brazil_ L2 - https://bmcgastroenterol.biomedcentral.com/articles/10.1186/1471-230X-12-49 DB - PRIME DP - Unbound Medicine ER -