Background

Polymerase chain reaction (PCR) technology and quantitative real-time PCR (qPCR) technology are widely used in clinical diagnosis and research, but amplification efficiency and sensitivity are still key problems for researchers. An increasing number of reports show that gold nanoparticles (AuNPs) can be used to improve the sensitivity and amplification efficiency of PCR. Here, we found that 60 nm gold nanoparticles with a positive charge (60 nm- Au⁺) can enhance the amplification efficiency of qPCR.

Methods

Mouse DNA was extracted by the alkaline lysis method. Primer 5.0 software was used to design primers and mutation primers, and the DNA fragments were obtained by the method of synthesizing plasmids. The qPCR was applied to amplify target gene fragments.

Results

The amplification efficiency of qPCR was improved by about 1.828 times in the experimental group with 60 nm- Au⁺ compared with the control group without 60 nm- Au⁺. The primer pair contains a specific palindromic sequence (GGATCC or ACCGGT). And 60 nm Au⁺ did not enhance the amplification efficiency of qPCR when the above primer was mutated.

Conclusions

The primers contain special palindrome sequences (GGATCC or ACCGGT) with 60 nm- Au⁺ can enhance the amplification efficiency of qPCR. Therefore, it suggests a more in-depth understanding of the mechanism and function of gold nanoparticles and primer sequences. This study has presented some implications for gold nanoparticles application in the development of qPCR technology.

中文翻译：

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60 nm 金纳米粒子通过引物中的特定回文序列（GGATCC 或 ACCGGT）提高 qPCR 扩增效率

背景

聚合酶链式反应（PCR）技术和实时定量PCR（qPCR）技术广泛应用于临床诊断和研究，但扩增效率和灵敏度仍然是研究人员面临的关键问题。越来越多的报告表明，金纳米粒子（AuNPs）可用于提高PCR的灵敏度和扩增效率。在这里，我们发现带正电荷的60 nm金纳米粒子（60 nm- Au ⁺）可以提高qPCR的扩增效率。

方法

采用碱裂解法提取小鼠DNA。采用Primer 5.0软件设计引物和突变引物，通过合成质粒的方法获得DNA片段。qPCR 用于扩增目标基因片段。

结果

^{与未加60 nm-Au +}的对照组相比，加60 nm-Au ⁺的实验组qPCR扩增效率提高约1.828倍。引物对包含特定的回文序列（GGATCC 或 ACCGGT）。当上述引物发生突变时， 60 nm Au ⁺并没有增强 qPCR 的扩增效率。

结论

引物含有60 nm-Au ⁺的特殊回文序列（GGATCC或ACCGGT） ，可以增强qPCR的扩增效率。因此，建议更深入地了解金纳米颗粒和引物序列的机制和功能。这项研究对金纳米粒子在 qPCR 技术开发中的应用提出了一些启示。