Genotyping of Single Nucleotide Polymorphisms (SNPs) Influencing Drug Response by Competitive Allele-specific Short Oligonucleotide Hybridization (CASSOH) with Immunochromatographic Strip

Hiratsuka, Masahiro; Ebisawa, Aiko; Matsubara, Yoichi; Kure, Shigeo; Konno, Yumiko; Sasaki, Takuma; Mizugaki, Michinao

doi:10.2133/dmpk.19.303

Cited by 8 publications

(3 citation statements)

References 24 publications

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“…65 AuNPs can be surface-modified with the analyte targeting components by surface passivation using sulphur-linked (for example, thiols and thioctic acids) 66 or electrostatically charged antibodies or DNA strands for example. 39 Capping ligands include; zwitterions, polymers, peptides, proteins, sugars and nucleic acids (Fig. 3).…”

Section: The Components Of Lateral Flowmentioning

confidence: 99%

“…38 Strategies to immobilise polynucleotides have also been reported. 39,40 The low absorption of unconjugated antibodies was noted in polymeric systems by Aoyama et al 41 To overcome this they produced high surface area microcone architectures on polycarbonate sheets. The increased surface area enabled high antibody immobilisation levels.…”

Section: The Components Of Lateral Flowmentioning

confidence: 99%

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Glycosylated gold nanoparticles in point of care diagnostics: from aggregation to lateral flow

et al. 2022

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Section: The Components Of Lateral Flowmentioning

confidence: 99%

Section: The Components Of Lateral Flowmentioning

confidence: 99%

Glycosylated gold nanoparticles in point of care diagnostics: from aggregation to lateral flow

et al. 2022

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“…The competitive allele-specific short oligonucleotide hybridization assay (CASSOH) with immunochromatographic strip 10,11 embraces the principle of the ASO hybridization method for allele discrimination in a dipstick-type assay format. Recently, we reported a dryreagent dipstick assay for SNP genotyping based on the PEXT reaction.…”

Section: Introductionmentioning

confidence: 99%

Dual-allele dipstick assay for genotyping single nucleotide polymorphisms by primer extension reaction

2008

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We have developed a dry-reagent dipstick test for simultaneous visual detection of two alleles in single nucleotide polymorphisms (SNPs). The strip comprises two test zones and a control zone. Oligonucleotidefunctionalized gold nanoparticles are used as reporters. PCR-amplified DNA that spans the interrogated sequence is subjected to primer extension (PEXT) reactions using allele-specific primers. Digoxigenin-dUTP and biotin-dUTP are incorporated in the extended fragments. The primers contain an oligo(dA) segment at the 5 0 end. The PEXT products are applied to the sample area of the strip, which is then immersed in the appropriate buffer. As the buffer migrates along the strip by capillary action, the extension products of the two alleles are captured at the test zones from immobilized anti-digoxigenin and streptavidin, whereas the oligo(dA) segment of the primers hybridizes with oligo(dT) strands attached to gold nanoparticles, thus generating characteristic red lines. The excess nanoparticles are captured from immobilized oligo(dA) strands at the control zone of the strip. The test was applied to the genotyping of two SNPs of the Toll-like receptor 4 gene (Asp299Gly and Thr399Ile), one SNP of CYP2C19 gene (CYP2C19*3) and one SNP of the TPMT gene (TPMT*2). Contrary to most genotyping methods, the dipstick test does not require costly specialized equipment for detection of PEXT products. The PCR product is pipetted directly into the PEXT reaction mixture without prior purification. The high sensitivity of the strip allows completion of PEXT reaction in three cycles only (7 min). The visual detection of both alleles is complete in 15 min.

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A novel enhancement assay for immunochromatographic test strips using gold nanoparticles

et al. 2006

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The immunochromatographic assay is a well-known and convenient diagnostic system. In this report, the development of a novel enhancement assay for the test strips is described. Additionally, this highly sensitive immunochromatographic assay was applied to detect human chorionic gonadotropin hormone (HCG) as the model case. The primary antibody-conjugated gold nanoparticles were used as the enhancer of the standard method. The primary antibodies were immobilized within a defined detection zone (test line) on the diagnostic nitrocellulose membrane. The secondary antibodies were conjugated with colloidal gold nanoparticles. In combination with an effective sample pretreatment, the gold-conjugated antibodies and the primary antibodies formed a sandwich complex with the target protein. Within the test line, the sandwich complex was immobilized, and furthermore, concentrated by the enhancer resulting in a localized surface plasmon resonance (LSPR) phenomenon and a distinct red color on the test line. The intensity of color of the red test line (signal intensity), which correlated directly with the concentration of the target protein in the standard or spiked samples, was assessed visually and by computer image analysis using a three-determination analysis. Under optimum conditions, the limit of detection (LOD) for HCG assay was 1 pg/mL. When using human serum, 10 pg/mL of HCG could be detected. We have also spiked total prostate-specific antigen (TPSA) in female serum. The LOD for TPSA was determined as 0.2 ng/mL. With this method, the quantitative determination of the target protein could be completed in less than 15 min. Our novel immunochromatographic strips using the enhancing method based on LSPR of gold nanoparticles are useful as a rapid and simple screening method for the detection of important analytes for medical applications, environmental monitoring, food control, and biosecurity.

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Genotyping of Single Nucleotide Polymorphisms (SNPs) Influencing Drug Response by Competitive Allele-specific Short Oligonucleotide Hybridization (CASSOH) with Immunochromatographic Strip

Cited by 8 publications

References 24 publications

Glycosylated gold nanoparticles in point of care diagnostics: from aggregation to lateral flow

Glycosylated gold nanoparticles in point of care diagnostics: from aggregation to lateral flow

Dual-allele dipstick assay for genotyping single nucleotide polymorphisms by primer extension reaction

A novel enhancement assay for immunochromatographic test strips using gold nanoparticles

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