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Abstract：
Acanthamoeba are widely distributed in the environment and are known to cause blinding keratitis and brain infections with greater than 90% mortality rate. Currently, polymerase chain reaction (PCR) is a highly sensitive and promising technique in Acanthamoeba detection. Remarkably, the rate of heating-cooling and convective heat transfer of the PCR tube is limited by low thermal conductivity of the reagents mixture. The addition of nanoparticles to the reaction has been an interesting approach that could augment the thermal conductivity of the mixture and subsequently enhance heat transfer through the PCR tube. Here, we have developed hexagonal boron nitride (hBN) nanoparticle-based PCR assay for the rapid detection of Acanthamoeba to amplify DNA from low amoeba cell density. As low as 1 × 10-4 wt % was determined as the optimum concentration of hBN nanoparticles, which increased Acanthamoeba DNA yield up to ~16%. Further, it was able to reduce PCR temperature that led to a ~2.0-fold increase in Acanthamoeba DNA yield at an improved PCR specificity at 46.2 °C low annealing temperature. hBN nanoparticles further reduced standard PCR step time by 10 min and cycles by eight; thus, enhancing Acanthamoeba detection rapidly. Enhancement of Acanthamoeba PCR DNA yield is possibly due to the high adsorption affinity of hBN nanoparticles to purine (Guanine-G) due to the higher thermal conductivity achieved in the PCR mixture due to the addition of hBN. Although further research is needed to demonstrate these findings in clinical application, we propose that the interfacial layers, Brownian motion, and percolation network contribute to the enhanced thermal conductivity effect.
摘要：
棘阿米巴在环境中广泛分布，已知会导致致盲性角膜炎和脑部感染，死亡率超过90％。目前,聚合酶链反应（PCR）是一种高度敏感且有前途的棘阿米巴检测技术。值得注意的是,例如,PCR管的加热-冷却和对流传热的速率受到试剂混合物的低热导率的限制。向反应中添加纳米颗粒是一种有趣的方法，可以增加混合物的热导率，并随后增强通过PCR管的传热。这里,我们已经开发了基于六方氮化硼（hBN）纳米颗粒的PCR测定法，用于快速检测棘阿米巴，以从低变形虫细胞密度扩增DNA。低至1×10-4重量%被确定为hBN纳米粒子的最佳浓度，这将棘阿米巴DNA产量提高到~16%。Further,它能够降低PCR温度，从而在46.2°C的低退火温度下以提高的PCR特异性使棘阿米巴DNA产量增加〜2.0倍。hBN纳米颗粒进一步将标准PCR步骤时间减少了10分钟，循环时间减少了8分钟；因此，快速增强棘阿米巴检测。棘阿米巴PCRDNA产量的提高可能是由于hBN纳米颗粒对嘌呤（鸟嘌呤-G）的高吸附亲和力，这是由于添加hBN在PCR混合物中获得了更高的热导率。尽管需要进一步的研究来证明这些发现在临床应用中，我们建议界面层，布朗运动,和渗滤网络有助于增强导热效应。
