Determination of mutations of the 23S rRNA gene of <i>Helicobacter pylori</i> by allele specific primer‐polymerase chain reaction method

Nakamura, Akiko; Furuta, Takahisa; Shirai, Naohito; Sugimoto, Mitsushige; Kajimura, Masayoshi; Soya, Yoshihiro; Hishida, Akira

doi:10.1111/j.1440-1746.2006.04546.x

Cited by 30 publications

(32 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…On the same culture media, mutant cells would be surpassed by susceptible cells, producing a misleading bacterial inoculum with a higher ratio of the wild genotype. In such situations, only an appropriate molecular method can discern a few H. pylori cells carrying resistant alleles (11,13) . Gastric samples showing H. pylori with resistant genotypes, 21 (77.8%) were also infected with strains carrying the wildtype allele, a high number of biopsies with mixed populations in comparison with other studies (11,12,13,17,20) .…”

Section: Resultsmentioning

confidence: 99%

“…The wild-type gene could be amplified alone in two of the three vessels (i.e., without competition from an occasional resistant allele). When DNA templates derived from resistant and susceptible H. pylori strains are simultaneously assayed by PCR in the same vessel, the under-represented allele would have its amplification partly harmed (11,17) . PCR formats, based on analysis of post-amplification products (e.g., polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and probe hybridizations followed by thermal melting analysis) would have difficulty detecting wild-type and mutant alleles.…”

Section: Resultsmentioning

confidence: 99%

“…PCR formats, based on analysis of post-amplification products (e.g., polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and probe hybridizations followed by thermal melting analysis) would have difficulty detecting wild-type and mutant alleles. A recently developed PCR technique, allelespecific primer-PCR (11) , can overcome occasional amplification competition between resistant and susceptible genotypes. In this format, specific primers, each one corresponding to wild type and mutant alleles, are substituted for probes, and each allele is amplified separately.…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Clarithromycin-resistant Helicobacter pylori in Recife, Brazil, directly identified from gastric biopsies by polymerase chain reaction

Lins

Lima

Magalhães

2010

Arq. Gastroenterol.

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Clarithromycin-resistant Helicobacter pylori in Recife, Brazil, directly identified from gastric biopsies by polymerase chain reaction

Lins

Lima

Magalhães

2010

Arq. Gastroenterol.

View full text Add to dashboard Cite

show abstract

“…CAM resistance has been found to result from a single-nucleotide substitution on the 23S rRNA gene of HP (2142A>G or C, 2143A>G) and CAM resistance can be predicted by detection of this nucleotide substitution. CAM resistance has been analyzed by the CAM susceptibility test using cultured HP and by the PCR-RFLP method or the invader method using HP DNA, but these methods pose problems related to rapidity, operability, economy and universality (8)(9)(10)(11)(12)(13)(14)24).…”

Section: Introductionmentioning

confidence: 99%

Personalized treatment in the eradication therapy for Helicobacter pylori

Jinda

Nakatani

Nishioka³

et al. 2011

Int J Mol Med

View full text Add to dashboard Cite

Abstract.Helicobacter pylori (HP) is known to be a causative bacterium of gastritis and peptic ulcers. The combination treatment consisting of a proton pump inhibitor (PPI), amoxicillin and clarithromycin (CAM) is widely used in eradication therapy, but the eradication fails in some patients. The main causes are CAM resistance of HP and individual variability in PPI metabolism related to the activity of the cytochrome P450 2C19 (CYP2C19) enzyme. In this study, we examined the usefulness of the prediction of the pharmacotherapeutic efficacy using a newly developed analysis system for HP CAM resistance and CYP2C19 genotypes. After obtaining the informed consent from 45 subjects with HPpositive peptic ulcers, biopsy specimens of the gastric mucosa were obtained by endoscopy. HP DNA extracted from the gastric mucosa was examined by the SELMAP-PCR method, the direct sequencing method or the singlenucleotide primer extension (SNuPE) method. HP detection rates by culture and the SELMAP-PCR method were 71% and 100%, respectively. Among 32 cultured HP, CAM resistance was confirmed in 6 samples by the in vitro drug susceptibility test. CAM-resistant gene mutations were also examined by the SELMAP-PCR method using 32 DNAs from cultured HP and the results were consistent with the drug susceptibility test. Among 22 patients, the eradication rate was 77%. Among 4 patients with CAM resistance determined by both the in vitro drug susceptibility test and the SNuPE method, eradication was successful in one intermediate metabolizer (IM), but not in three extensive metabolizers (EMs). Patients were divided into three groups according to their CYP2C19 phenotype: EMs, IMs and poor metabolizers (PMs). The eradication rates for 6 EMs, 12 IMs and 4 PMs were 33.3%, 91.7% and 100%, respectively. Based on these results, the information on CAM resistance in HP and CYP2C19 phenotypes in carriers could predict the pharmacotherapeutic efficacy and probability of eradication. It can then be possible to vary the dosing or to select another drug by the prediction of the pharmacotherapeutic efficacy.

show abstract

“…The allele-specific primer polymerase chain reaction (ASP-PCR) assay is a method used to determine SNPs based on DNA amplification, and the SNPs can be determined by whether the primers anneal to the target DNA [17,18] . The present study was designed to determine SNPs at position 808 of the SLC22A2 gene using ASP-PCR and to investigate the potential relationship between SLC22A2 gene polymorphisms and blood lactate concentrations in Shanghai Hans with T2DM.…”

Section: Introductionmentioning

confidence: 99%

SLC22A2 gene 808 G/T variant is related to plasma lactate concentration in Chinese type 2 diabetics treated with metformin

Liu

Zheng

et al. 2010

Acta Pharmacol Sin

View full text Add to dashboard Cite

Aim: To investigate the potential relationship between the SLC22A2 gene polymorphism and blood lactate concentration in Shanghai Hans suffering from type 2 diabetes mellitus (T2DM). Methods: The SLC22A2 single nucleotide polymorphism (SNP) 808G/T was genotyped in 400 T2DM patients, including a metformintreated group (n=200) and a non-metformin-treated group (n=200). Fasting plasma lactic acid levels were measured with an enzymeelectrode assay. Biochemical indexes, including plasma alanine aminotransferase (ALT), creatinine (Cr), and glycolated hemoglobin (HbA1c), were also measured. Results: The fasting plasma lactate concentration in the metformin-treated group was significantly higher than that in the non-metformin-treated group (1.29±0.45 mmol/L vs 1.18±0.44 mmol/L, P=0.015). Additionally, the ratio of patients with hyperlactacidemia was 8% (16/200) for the metformin-treated group and 5.5% (11/200) for the non-metformin-treated group, with no lactic acidosis found in either group. The frequency of the SLC22A2 808G/T T allele was 12.9%. Patients with the mutant genotype (TT) had a higher blood lactate concentration in the metformin-treated group than those in the non-metformin-treated group (t=2.492, P=0.013). This trend was not observed in the GG and GT genotypes when compared with metformin-treated and non-metformin-treated groups. Patients with the mutant genotype (TT) in the metformin-treated group also had a higher incidence of hyperlactacidemia compared with the GG genotype (40.0% vs 6.9%, P=0.050) in the metformin-treated group and the GG (6.0%, P=0.042) or GT (4.3%, P=0.043) genotypes in the non-metformin-treated group. In the metformin-treated group, there were significant gender differences in lactate concentrations in the TT (2.18±0.15 vs 1.04±0.27 mmol/L, P=0.008) and GG genotypes (1.40±0.51 vs 1.19±0.35 mmol/L, P=0.004). The lactate levels of women with the TT genotype were the highest in the metformin-treated group, but differences in lactate levels among the genotypes were not observed in the non-metformin-treated group. Conclusion: There is an 808G/T polymorphism in the SLC22A2 gene in Chinese Hans with T2DM. The 808G>T variance in the SLC22A2 gene can affect the plasma lactate level and the incidence of hyperlactacidemia in T2DM patients undergoing metformin therapy. Additionally, the female patients carrying the TT genotype are prone to lactatemia.

show abstract

Determination of mutations of the 23S rRNA gene of Helicobacter pylori by allele specific primer‐polymerase chain reaction method

Abstract: The ASP-PCR assay for 23S rRNA mutation of H. pylori is a useful method to detect clarithromycin-resistant strains of H. pylori easily.

Cited by 30 publications

References 39 publications

Clarithromycin-resistant Helicobacter pylori in Recife, Brazil, directly identified from gastric biopsies by polymerase chain reaction

Clarithromycin-resistant Helicobacter pylori in Recife, Brazil, directly identified from gastric biopsies by polymerase chain reaction

Personalized treatment in the eradication therapy for Helicobacter pylori

SLC22A2 gene 808 G/T variant is related to plasma lactate concentration in Chinese type 2 diabetics treated with metformin

Contact Info

Product

Resources

About