A Simple and Rapid Genotyping Assay for Simultaneous Detection of Two ADRB2 Allelic Variants Using Fluorescence Resonance Energy Transfer Probes and Melting Curve Analysis

Sabato, Mariangela; Irani, Anne-Marie; Bukaveckas, Bonny L.; Schwartz, Lawrence B.; Wilkinson, David S.; Ferreira‐Gonzalez, Andrea

doi:10.2353/jmoldx.2008.070133

Cited by 5 publications

(3 citation statements)

References 41 publications

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“…The target DNA sequence of the B2ADR NM 000024.4 was amplified using a set of primers that were previously described 25,26 (forward, nucleotides 188---212: 5 -AGCCAGTGCGCTCAC-CTGCCAGACT-3 ; reverse, nucleotides 406---383: 5 -GCTCGAACTTGGCAATGGC-TGTGA-3 ) to generate an amplicon of 219 bp in length. Allelic discrimination assay for SNPs relating to the B2ADR expression (rs 1042713) was carried out using previously described SNPs detective system, sequence-specific thermal-elution chromatography.…”

Section: ß2-adrenergic Receptor (B2adr) Genes Analysismentioning

confidence: 99%

Recent advance in investigation of gene polymorphisms in Japanese patients with aspirin-exacerbated respiratory disease

Kurosawa¹,

Yukawa²,

Hozawa³

et al. 2015

Allergologia et Immunopathologia

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Aspirin-exacerbated respiratory disease (AERD) is a complex clinical syndrome characterised by severe asthmatic attack upon treatment with aspirin and/or non-steroidal anti-inflammatory drugs (NSAIDs). Genetic predisposition has been considered as a crucial determinant and candidate genes have concentrated especially on cysteinyl leukotrienes (LTs)-related genes as the inhibitory action of aspirin and NSAIDs on cyclooxygenase activity may cause overproduction of cysteinyl LTs. However, conflicting results have been reported, in parallel with replication studies in different ethnic groups. Thus, future areas of investigations need to focus on comprehensive approaches towards the discovery of other genetic biomarkers. Unfortunately, few papers have been reported about gene polymorphisms in Japanese patients with AERD. Here, we described on our recent genetic investigations on B2ADR, IL-13, IL-17A, CYP2C19, TBXA2R, CRTH2 and HSP70. This review indicates potential genetic biomarkers contributing to the early diagnosis of AERD, which may include CYP2C19 and HSP70 gene polymorphisms, and future validation studies in independent population are required to provide reassurance about our findings.

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Section: ß2-adrenergic Receptor (B2adr) Genes Analysismentioning

confidence: 99%

Recent advance in investigation of gene polymorphisms in Japanese patients with aspirin-exacerbated respiratory disease

Kurosawa¹,

Yukawa²,

Hozawa³

et al. 2015

Allergologia et Immunopathologia

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“…DNA in the specimens obtained by rubbing buccal mucosa with a cotton swab from the patients was extracted by using QIAamp 96 DNA blood kits (Qiagen, Hilden, Germany). The target DNA sequence of the ADRB2 gene NM_000024.4 was amplified using a set of primers that were previously described [15,16] (forward, nucleotides 188-212: 5'-AGCCAGTGCGCTCACCTGCCAGACT-3'; reverse, nucleotides 406-383:5'to generate an amplicon for single nucleotide expression (rs 1042713) sequence-specific thermal-elution chromatography our laboratory [17,18].…”

Section: Patients and Genotyping Of Adrb2 Polymorphismmentioning

confidence: 99%

A Pilot Study on Effect of Beta2-Adrenergic Receptor Gene Polymorphism on Response to Fluticasone/Formoterol Inhaler in Patients with Persistent Asthma

Yukawa¹

2015

J Allergy Ther

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Background: Safety concerns have been raised regarding the regular use of long-acting ß2-agonists (LABAs) with inhaled corticosteroids (ICSs), and some studies suggested patients with asthma who are homozygous for arginine (ArgArg) at the 16th amino acid position of the ß2-adrenergic receptor (ADRB2) gene benefit less from treatment with ICS/LABA than do those homozygous for glycine (GlyGly). Phase III trial of fluticasone/formoterol, which consisted of 8-week, multicenter randomized single-blinded parallel-group study and 52-week, multicenter open-label study, have been conducted in Japan. In the trials, we performed a pilot study on effect of ADRB2 gene polymorphisms on lung function and asthma control to regular use of the new ICS/LABA inhaler. Methods: As number of patients was limited to each institution, nine patients were allowed to participate in this pilot study. DNA in the specimens in patients with persistent asthma was extracted, and the target DNA sequence of the gene was amplified. Allelic discrimination assay for single nucleotide polymorphisms relating to the ADRB2 was carried out. Results: In the 8-week study, patients with either ArgArg or ArgGly genotype showed good changes of peak expiratory flow (PEF) values from baseline with fluticasone/formoterol treatment. In the 52-week study, the improvement of lung function tests (forced expiratory volume in one second and PEF) from baseline was observed with the treatment, and all of the daily diary-based measures of asthma control were improved from baseline in each patient over the treatment period irrespective of their genotypes of ADRB2. Conclusion: This is the first prospective pilot study, which might suggest inheritance of ADRB2 gene polymorphisms may not appear to effect clinical outcomes with fluticasone/formoterol inhaler.

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“…The target DNA sequence of the ADRB2 NM_000024.4 was amplified using a set of primers that were previously described [35,36] (forward, nucleotides 188–212: 5′-AGCCAGTGCGCTCACCTGCCAGACT-3′; reverse, nucleotides 406–383: 5′-GCTCGAACTTGGCAATGGCTGTGA-3′) to generate an amplicon of 219 bp in length. Allelic discrimination assay for SNPs relating to the ADRB2 expression (rs 1042713) was carried out by using a previously described SNPs detective system, sequence-specific thermal-elution chromatography [37].…”

Section: Methodsmentioning

confidence: 99%

Arg16Gly β2-Adrenergic Receptor Gene Polymorphism in Japanese Patients with Aspirin-Exacerbated Respiratory Disease

Kohyama¹,

Abe²,

Kodaira³

et al. 2011

Int Arch Allergy Immunol

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Background: There has been no report that investigated β₂-adrenergic receptor (ADRB2) gene polymorphism in patients with aspirin-exacerbated respiratory disease (AERD). Methods: DNA in the specimens in three groups of study subjects classified patients with AERD, patients with aspirin-tolerant asthma (ATA) and normal controls was extracted, and the target DNA sequence of the ADRB2 was amplified using a set of primers to generate an amplicon of 219 bp in length. Allelic discrimination assay for single nucleotide polymorphisms relating to the ADRB2 gene expression was carried out by using a previously described single nucleotide polymorphism detective system, sequence-specific thermal-elution chromatography. Results: The frequency of the Gly variant allele in patients with AERD was significantly lower than that in patients with ATA (p = 0.007), and the odds ratio (OR) of AERD to ATA associated with wild-type ArgArg homozygote was 3.300. Frequencies of wild-type ArgArg homozygote are significantly higher than those of variant-type ArgGly/GlyGly genotype in patients with AERD compared with those with ATA (p < 0.001, OR = 3.153). In patients with AERD, frequencies of wild-type ArgArg homozygote in both female and male patients are significantly higher than those of variant-type ArgGly/GlyGly genotype in male patients compared with those with ATA (p < 0.001, OR = 5.128 and p = 0.007, OR = 4.367, respectively). Also, in patients with AERD, frequencies of wild-type ArgArg homozygote in female patients are significantly higher than those of variant-type ArgGly/GlyGly genotype in female patients compared with those with ATA (p = 0.002, OR = 2.825). Conclusions: We were the first to analyze Arg16Gly ADRB2 gene polymorphism in Japanese patients with AERD, and showed that Arg16Gly ADRB2 gene polymorphism in Japanese patients with AERD is different from that in the patients with ATA.

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A Simple and Rapid Genotyping Assay for Simultaneous Detection of Two ADRB2 Allelic Variants Using Fluorescence Resonance Energy Transfer Probes and Melting Curve Analysis

Cited by 5 publications

References 41 publications

Recent advance in investigation of gene polymorphisms in Japanese patients with aspirin-exacerbated respiratory disease

Recent advance in investigation of gene polymorphisms in Japanese patients with aspirin-exacerbated respiratory disease

A Pilot Study on Effect of Beta2-Adrenergic Receptor Gene Polymorphism on Response to Fluticasone/Formoterol Inhaler in Patients with Persistent Asthma

Arg16Gly β2-Adrenergic Receptor Gene Polymorphism in Japanese Patients with Aspirin-Exacerbated Respiratory Disease

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