Upadhyay Archana, Yang Huan, Zaman Bilal, Zhang Lei, Wu Yundi, Wang Jinhua, Zhao Jianguo, Liao Chenghong, Han Qian
Laboratory of Tropical Veterinary Medicine and Vector Biology, School of Life and Pharmaceutical Sciences, Hainan University, Haikou, Hainan 570228, China.
State Key Laboratory of Marine Resource Utilization in South China Sea, College of Material Science and Engineering, Haikou, Hainan 570228, China.
Pathogens. 2020 Feb 14;9(2):122. doi: 10.3390/pathogens9020122.
Polymerase chain reaction (PCR) is a unique technique in molecular biology and biotechnology for amplifying target DNA strands, and is also considered as a gold standard for the diagnosis of many canine diseases as well as many other infectious diseases. However, PCR still faces many challenges and issues related to its sensitivity, specificity, efficiency, and turnaround time. To address these issues, we described the use of unique ZnO nanoflowers in PCR reaction and an efficient ZnO nanoflower-based PCR (nanoPCR) for the molecular diagnosis of canine vector-borne diseases (CVBDs). A total of 1 mM of an aqueous solution of ZnO nanoflowers incorporated in PCR showed a significant enhancement of the PCR assay with respect to its sensitivity and specificity for the diagnosis of two important CVBDs, and . Interestingly, it drastically reduced the turnaround time of the PCR assay without compromising the yield of the amplified DNA, which can be of benefit for veterinary practitioners for the improved management of diseases. This can be attributed to the favorable adsorption of ZnO nanoflowers to the DNA and thermal conductivity of ZnO nanoflowers. The unique ZnO nanoflower-assisted nanoPCR greatly improved the yield, purity, and quality of the amplified products, but the mechanism behind these properties and the effects and changes due to the different concentrations of ZnO nanoflowers in the PCR system needs to be further studied.
聚合酶链反应(PCR)是分子生物学和生物技术领域中用于扩增目标DNA链的一项独特技术,同时也被视为许多犬类疾病以及许多其他传染病诊断的金标准。然而,PCR在灵敏度、特异性、效率和周转时间方面仍面临诸多挑战和问题。为解决这些问题,我们描述了在PCR反应中使用独特的氧化锌纳米花以及一种基于氧化锌纳米花的高效PCR(纳米PCR)用于犬媒传播疾病(CVBDs)的分子诊断。在PCR中加入总共1 mM的氧化锌纳米花水溶液,对于两种重要的CVBDs([此处原文缺失两种疾病名称])的诊断,其PCR检测在灵敏度和特异性方面均有显著提高。有趣的是,它大幅缩短了PCR检测的周转时间,同时不影响扩增DNA的产量,这对兽医改善疾病管理有益。这可归因于氧化锌纳米花对DNA的良好吸附以及氧化锌纳米花的热导率。独特的氧化锌纳米花辅助纳米PCR极大地提高了扩增产物的产量、纯度和质量,但这些特性背后的机制以及PCR系统中不同浓度氧化锌纳米花所产生的影响和变化仍需进一步研究。