Nanomaterials-based Polymerase Chain Reactions for DNA Detection

Pan, Dun; Wen, Yangqin; Mi, Lijuan; Fan, Chunhai; Hu, Jun

doi:10.2174/138527211794474447

Cited by 22 publications

(14 citation statements)

References 77 publications

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“…Based on previous reports, the AuNP specificity enhancement of PCR might be caused by the nonspecific adsorption of AuNPs of single-strand DNA. 30 , 31 To assess this nonspecific absorption, the UV-Vis spectra of AuNPs and AuNPs mixed with NA sequences (including primers, probes, and single-strand templates) were measured ( Figure S3 ). A discernable redshift (~11 nm) in the surface plasmon resonance (SPR) peak of AuNPs was observed after mixing with these single-strand sequences (pink line in Figure S3 ).…”

Section: Resultsmentioning

confidence: 99%

Taqman-MGB nanoPCR for Highly Specific Detection of Single-Base Mutations

Xue¹,

You²,

Peng³

et al. 2021

IJN

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Purpose Detection of single-base mutations is important for real-time monitoring of tumor progression, therapeutic effects, and drug resistance. However, the specific detection of single-base mutations from excessive wild-type background sequences with routine PCR technology remains challenging. Our objective is to develop a simple and highly specific qPCR-based single-base mutation detection method. Methods Using EGRF T790M as a model, gold nanoparticles at different concentrations were separately added into the Taqman-MGB qPCR system to test specificity improvement, leading to the development of the optimal Taqman-MGB nanoPCR system. Then, these optimal conditions were used to test the range of improvement in the specificity of mutant-type and wild-type templates and the detection limit of mutation abundances in a spiked sample. Results The Taqman-MGB nanoPCR was established based on the traditional qPCR, with significantly suppressed background noise and improved specificity for single-base mutation detection. With EGFR T790M as a template, we demonstrated that our Taqman-MGB nanoPCR system could improve specificity across a wide concentration range from 10 −9 μM to 10 μM and detect as low as 0.95% mutation abundance in spiked samples, which is lower than what the traditional Taqman-MGB qPCR and existing PCR methods can detect. Moreover, we also proposed an experimentally validated barrier hypothesis for the mechanism of improved specificity. Conclusion The developed Taqman-MGB nanoPCR system could be a powerful tool for clinical single-base mutation detection.

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Section: Resultsmentioning

confidence: 99%

Taqman-MGB nanoPCR for Highly Specific Detection of Single-Base Mutations

Xue¹,

You²,

Peng³

et al. 2021

IJN

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“…It was found that the optimal GNPs concentration in the PCR reaction was 1 nM, which had a 3-fold higher band intensity compared to a reaction without the addition of any GNPs (Fig. 29,30 The inhibition effect can occur with high concentration of GNPs where DNA polymerase can be strongly but not completely adsorbed on the negatively charged surface of GNPs, with their polar groups modulating the amount of active polymerases in the reaction. The intensity of the band increases as the concentration of the GNPs increases up to 1 nM, then it starts to decline with higher GNPs concentrations such as 1.5 and 2 nM.…”

Section: Effect Of Different Gnps Concentrations On Conventional Pcrmentioning

confidence: 97%

The effect of gold nanoparticles on the diagnostic polymerase chain reaction technique for equine herpes virus 1 (EHV-1)

2016

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Nano-biotechnology has been a noticeable research area because of its successful applications in molecular diagnostics and therapy of various genetic and microbial diseases. Although the polymerase chain reaction (PCR) technique is one of the most highlighted and promising applications in the molecular diagnosis field, it suffers from some drawbacks that affect its efficiency. For instance, as a diagnostic technique for equine herpes virus-1 (EHV-1), conventional PCR could lead to false negative results due to the low viral titer in some samples, which leads to the necessity to improve its sensitivity.In this study, we carried out experiments to determine the effects of 15 nm unmodified citrate-coated gold nanoparticles (GNPs) on the key PCR reactants in order to see if these would enhance the overall outcomes of the reaction. Our results showed that, after optimization of the GNPs, oligonucleotide primers and Taq polymerase concentrations, a specific high yield amplification with a detection limit of 10 2 DNA copies could be reached compared to the 10 5 to 10 4 detection limit of conventional PCR. Thus, the developed and optimized GNPs-assisted PCR technique could be used for a more efficient, highly sensitive molecular detection of EHV-1.

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“…Nanoparticles through their properties in efficiency of heat transfer can work as modulators to the PCR (Khaliq et al, 2010), and is reported that metal nanoparticles (Au) and oxides nanoparticles (TiO 2 ) have been commonly used in improve PCR efficiency since the first report of nanoparticleassisted PCR was published (Pan et al, 2011), so that the metal nanoparticles can enhance PCR specificity by applying the appropriate concentration and provide better bands of the target product. These properties are extremely useful in the case of Lxx detection in plant tissue, since most of the inconsistencies in the method are related to nonoccurrence of the reaction, observed as no bands on the running gel, or when the PCR is effective but non-specific, leading to the formation of nonspecific bands.…”

Section: Nano Pcr Analysismentioning

confidence: 99%

Nanoparticle-assisted Polymerase Chain Reaction (NanoPCR): Optimization of PCR detection of Leifsonia xyli subsp. xyli by the addition of nanoparticles

Dantas

Melo

Houllou

et al. 2017

jaar

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Leifsonia xyli subsp. xyli (Lxx) causes ratoon stunting disease (RSD) in sugarcane, and is one of major causes of production losses. The detection of Lxx bacteria in sugarcane is made mainly through molecular biology techniques, especially polymerase chain reaction (PCR). However, PCR presents some barriers to provide reliable results. The present work brings a Nanoparticle-assisted Polymerase Chain Reaction (NanoPCR) assay for the detection of Lxx in its latent infection on micropropagated sugarcane. This assay was based in the addition of Gold and Titanium dioxide nanoparticles to conventional PCR and evaluation of its effects. It was observed that the reactions performed with Titanium dioxide nanoparticles provided the formation of singular well-defined bands under electrophoresis, consistent with the expected molecular weight, without occurrence of non-specific bands or presenting false negatives occurrence, negative effects that were observed in the control assay. While the performed NanoPCR adding AuNP also provided the formation of well-defined bands, been able to inhibit the occurrence of false negatives, but wasn't able to eliminate the occurrence of non-specific amplifications. The results indicate that NanoPCR by the addition of Gold and Titanium dioxide nanoparticles to conventional PCR increased the detection of Lxx.

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Nanomaterials-based Polymerase Chain Reactions for DNA Detection

Cited by 22 publications

References 77 publications

Taqman-MGB nanoPCR for Highly Specific Detection of Single-Base Mutations

Taqman-MGB nanoPCR for Highly Specific Detection of Single-Base Mutations

The effect of gold nanoparticles on the diagnostic polymerase chain reaction technique for equine herpes virus 1 (EHV-1)

Nanoparticle-assisted Polymerase Chain Reaction (NanoPCR): Optimization of PCR detection of Leifsonia xyli subsp. xyli by the addition of nanoparticles

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