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用于检测的六方氮化硼纳米颗粒辅助快速热循环

hBN Nanoparticle-Assisted Rapid Thermal Cycling for the Detection of .

作者信息

Rasheed Abdul Khaliq, Siddiqui Ruqaiyyah, Ahmed Salma Mohammed Kabir, Gabriel Shobana, Jalal Mohammed Zayan, John Akbar, Khan Naveed Ahmed

机构信息

Department of New Energy Science and Engineering, School of Energy and Chemical Engineering, Xiamen University Malaysia Campus, Bandar Sunsuria, Sepang, Selangor 43900, Malaysia.

Department of Biology, Chemistry and Environmental Sciences, College of Arts and Sciences, American University of Sharjah, Sharjah 26666, UAE.

出版信息

Pathogens. 2020 Oct 7;9(10):824. doi: 10.3390/pathogens9100824.

DOI:10.3390/pathogens9100824
PMID:33036480
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7601326/
Abstract

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 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 to amplify DNA from low amoeba cell density. As low as 1 × 10 wt % was determined as the optimum concentration of hBN nanoparticles, which increased DNA yield up to ~16%. Further, it was able to reduce PCR temperature that led to a ~2.0-fold increase in 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 detection rapidly. Enhancement of 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

它们广泛分布于环境中,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)是一种检测中高度灵敏且有前景的技术。值得注意的是,PCR管的加热-冷却速率和对流热传递受试剂混合物低导热率的限制。向反应中添加纳米颗粒是一种有趣的方法,可提高混合物的导热率,进而增强通过PCR管的热传递。在此,我们开发了基于六方氮化硼(hBN)纳米颗粒的PCR检测方法,用于从低阿米巴细胞密度中快速检测并扩增DNA。确定低至1×10 wt%的hBN纳米颗粒浓度为最佳浓度,其使DNA产量提高了约16%。此外,它能够降低PCR温度,在46.2℃的低退火温度下,提高PCR特异性的同时使DNA产量提高约2.0倍。hBN纳米颗粒进一步将标准PCR步骤时间缩短了10分钟,循环次数减少了8次;从而快速增强检测效果。PCR DNA产量的提高可能是由于hBN纳米颗粒对嘌呤(鸟嘌呤-G)具有高吸附亲和力,这是由于添加hBN后PCR混合物实现了更高的导热率。尽管需要进一步研究以在临床应用中证实这些发现,但我们认为界面层、布朗运动和渗流网络有助于增强热导率效应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/ccaae1a0e5df/pathogens-09-00824-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/40378c886fd6/pathogens-09-00824-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/2325b7ce969a/pathogens-09-00824-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/61aeaad0e596/pathogens-09-00824-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/1ca467eaa9a1/pathogens-09-00824-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/d26806b69941/pathogens-09-00824-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/33b24fb6e3dd/pathogens-09-00824-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/56e3e8f34318/pathogens-09-00824-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/ccaae1a0e5df/pathogens-09-00824-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/40378c886fd6/pathogens-09-00824-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/2325b7ce969a/pathogens-09-00824-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/61aeaad0e596/pathogens-09-00824-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/1ca467eaa9a1/pathogens-09-00824-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/d26806b69941/pathogens-09-00824-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/33b24fb6e3dd/pathogens-09-00824-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/56e3e8f34318/pathogens-09-00824-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/004e/7601326/ccaae1a0e5df/pathogens-09-00824-g008.jpg

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